KR20080087820A - Docosahexaenoic acid (dha) producing thraustochytrid strain-sc1 - Google Patents

Abstract

The invention relates to the screening of an unreported strain of thraustochytrid-SC1 producing significant amounts of Docosahexaenoic acid (DHA) and also accumulate their respective intermediate fatty acids in addition to DHA. This organism has been isolated from the backwaters of Goa by Pollen Baiting method. The fatty acid profile and the 18S ribosomal sequences of each of this organism has been disclosed indicating its molecular phylogeny. The organism on further study can be very significant for their utilization in the commercial production of DHA.

Description

도코사헥사엔산〔ＤＨＡ〕을 생산하는 트라우스토키트리드 균주－ＳＣ１{DOCOSAHEXAENOIC ACID (DHA) PRODUCING THRAUSTOCHYTRID STRAIN-SC1}DOCOSAHEXAENOIC ACID (DHA) PRODUCING THRAUSTOCHYTRID STRAIN-SC1} Producing Docosahexaenoic Acid [DHA]

본 발명은 폴렌 미끼법(Pollen baiting method)에 의해 고아(Goa)의 역류로부터 분리된 상당량의 도코사헥사엔산을 생산하는 신규의 미공표된 트라우스토키트리드(Thraustochytrid) 균주-SC1의 선별(screening)에 관한 것이다. 상당량의 오메가-3-지방산의 생산을 나타내는 상기 분리된 유기체의 지방산 프로파일이 개시되어 있다. 또한, 그들의 분자 계통발생을 나타내는 분리된 유기체의 18S rRNA 서열도 밝히고 있다.The present invention provides for the selection of a novel, unpublished Thraustochytrid strain-SC1 that produces a significant amount of docosahexaenoic acid isolated from the reflux of Goa by the Pollen baiting method ( screening). Fatty acid profiles of the isolated organisms are described which represent the production of significant amounts of omega-3-fatty acid. It also reveals 18S rRNA sequences of isolated organisms that exhibit their molecular phylogeny.

지난 10년간 다불포화 지방산(PUFA: polyunsaturated fatty acid)의 영양상의 중요성에 대한 관심이 현저하게 증가되어 왔다. 다불포화 지방산은 2개 이상의 이중 결합을 함유하는 장쇄 지방산이다. 이들에 대한 관심은 치료, 식품 및 영양적 응용에 있어서의 그들의 잠재성에 기인한다. 이들은 선택된 종자 식물 및 몇몇 해양 공급원(marine source)으로부터 상업적으로 생산되고 있다. 그러나, 현재의 공급원으로부터 정제된 PUFA의 생산은 시장에의 공급에 적합하지 않게 될 것이다(Gill and Valivety 1997). 수요에 대한 기대 상승에 부응하고 어유(fish oil)의 결함을 타개하기 위해서, PUFA에 대한 대체 생산 프로세스가 현재 개발 중에 있 다.Over the last decade there has been a significant increase in the nutritional importance of polyunsaturated fatty acids (PUFAs). Polyunsaturated fatty acids are long chain fatty acids containing two or more double bonds. Their interest is due to their potential in therapeutic, food and nutritional applications. They are commercially produced from selected seed plants and several marine sources. However, the production of purified PUFAs from current sources will not be suitable for supply to the market (Gill and Valivety 1997). In order to meet rising expectations for demand and to address deficiencies in fish oil, alternative production processes for PUFA are currently under development.

PUFA는 지방산 사슬의 말단 탄소 원자로부터 3C 또는 6C가 되는 말단 결합의 위치를 토대로 해서 2개의 시리즈로 그룹화되어 있다. 이들은 일반적으로 2개의 주된 그룹, 즉, 오메가-6(ω6 혹은 n-6)과 오메가-3(ω3 혹은 n-3)으로 분류되고 있다. 오메가-6 지방산 중에서, 아라키돈산은 많은 프로스타글란딘류 및 에이코사노이드류의 주된 전구체이므로 특히 중요성이 증가하고 있고, 오메가-3 지방산 중에서, 도코사헥사엔산은 현재 많은 관심을 받고 있고 "필수 지방산"으로서 일컬어져 왔다.PUFAs are grouped into two series based on the position of the end bonds becoming 3C or 6C from the terminal carbon atoms of the fatty acid chain. These are generally classified into two main groups: omega-6 (ω6 or n-6) and omega-3 (ω3 or n-3). Among the omega-6 fatty acids, arachidonic acid is of particular importance because it is a major precursor of many prostaglandins and eicosanoids, and among the omega-3 fatty acids, docosahexaenoic acid is currently receiving much attention and is referred to as "essential fatty acid". come.

탄소수 22개의 장쇄의 불포화 오메가-3 지방산인 도코사헥사엔산(DHA: docosahexaenioic acid)은 어류 및 해양 식물에서 통상 발견되고 있다. 또한, 이것은 인간 뇌 조직의 주된 구성 블록이고 뇌의 회색질 및 망막에서의 일차 구조 지방산이라는 사실로 인해 주된 중요성을 얻고 있다. 최근의 지견들은 치매, 우울증, 기억 상실 및 인간의 DHA의 생리적 중요성을 입증하는 가시적인 문제점 등의 노화와 관련된 특정 거동 및 신경학적 병증 상태와 낮은 레벨의 DHA 간의 상관을 나타내고 있다.Docosahexaenioic acid (DHA), a long-chain unsaturated omega-3 fatty acid having 22 carbon atoms, is commonly found in fish and marine plants. It is also of primary importance due to the fact that it is the major building block of human brain tissue and is the primary structural fatty acid in the brain's gray matter and retina. Recent findings have shown a correlation between low levels of DHA and certain behavioral and neurological conditions related to aging, such as dementia, depression, memory loss and visible problems demonstrating the physiological importance of human DHA.

WHO는 최근 2000 칼로리 식사 섭취를 기준으로 매일 약 2.2 내지 4.4g에 상당하는 오메가-3 지방산으로부터 1일 섭취 에너지의 1 내지 2%를 커버하도록 권장하고 있다. 우리의 식사를 통한 오메가-3 PUFA의 섭취는 주로 해산물의 소비를 통해 일어나며, 이 해산물은 오메가-3 PUFA가 풍부한 것을 특징으로 하고 있다. 평균 섭취는 모집단 간에 다르지만, 이 강력한 수요는 대규모의 해양 어류 농장의 도 입을 가져왔고, 또한 수종의 해양 어류 유충의 정상 성장과 발달은 식사에서의 오메가-3 다불포화 지방산, 특히 DHA 및 에이코사펜타엔산의 보충에 의존한다(Rodriguez et al.1998).The WHO recently recommends 1 to 2% of the daily intake energy from about 2.2 to 4.4 grams of omega-3 fatty acids per day based on a 2000 calorie meal intake. The intake of omega-3 PUFAs through our meals is mainly due to the consumption of seafood, which is characterized by a rich in omega-3 PUFAs. While average intake varies among the populations, this strong demand has led to the introduction of large scale marine fish farms, and the normal growth and development of several marine fish larvae has led to omega-3 polyunsaturated fatty acids in the diet, especially DHA and eicosapenta. Rely on supplementation of enoic acid (Rodriguez et al. 1998).

현재 선택된 어유 및 미세조류 종은 DHA의 주된 산업적 공급원이다. 최고 레벨의 EPA 및 DHA를 가진 어유는 고등어, 청어 및 연어를 포함한다. 대구류(cod), 북대서양산 대구(haddock) 등의 몇몇 어류는 간에 대부분의 지방을 비축한다. 그럼에도 불구하고, 최상의 공급원은 예컨대 참치, 고등어, 정어리, 청어 및 송어 등의 냉수어이다. 그러나, 어유로부터 DHA의 최대의 이익을 얻기 위해서는, 사람들은 어류를 생으로 먹거나 끓여서 먹는 것이 바람직하고, 나아가서는 복부를 따라 그리고 지느러미 둘레의 아가미 뒤쪽의 껍질을 먹을 필요가 있으며, 그 이유는 이들 부분에는 오일의 대부분이 축적되어 있기 때문이다. 적절하게는, 빛, 열 및 산소가 모두 어유 중의 EPA 및 DHA를 감소시키는 것은 주지 사항이다. 어유는 고도로 다불포화되어 있기 때문에, 이들은 신속하게 부패한다. 부패한 어류는 비린내가 나므로 그다지 식욕을 돋구지 못한다. 어유로부터의 PUFA의 용도 혹은 유아용 혼합 분유에서의 그들의 내포도 많은 단점을 가진다. 어유는 일반적으로는 유아용 혼합 분유 중에 바람직하지 않은 성분인 에이코사펜탄산을 함유하는 데, 이것은 유아에게서 아라키돈산 레벨을 낮추기 때문이다. 이것은 유아 체중 증가의 감소 속도와 상관 관계를 가지고 있다.Currently selected fish oil and microalgal species are the major industrial sources of DHA. Fish oils with the highest levels of EPA and DHA include mackerel, herring and salmon. Some fish, such as cod and the north Atlantic haddock, stock most of the liver. Nevertheless, the best sources are cold water fish such as tuna, mackerel, sardines, herring and trout. However, in order to get the maximum benefit of DHA from fish oil, it is desirable for people to eat fish raw or boiled, and then to eat the shells along the abdomen and behind the gills around the fins, for these parts This is because most of the oil is accumulated. Suitably, it is well known that light, heat and oxygen all reduce EPA and DHA in fish oil. Because fish oils are highly polyunsaturated, they rot quickly. Corrupt fish are fishy, so they don't have much appetite. The use of PUFAs from fish oils or their inclusion in infant formulas also has many disadvantages. Fish oils generally contain eicosaptanic acid, an undesirable component in infant formulas, because it lowers arachidonic acid levels in infants. This correlates with the reduction rate of infant weight gain.

또, 어유의 공급사는 각종 수산업의 실패 혹은 가변성으로 인해 신뢰성이 없을 수 있다. 이것은 DHA의 세계적인 수요에 부응하도록 충분한 어유를 이용할 수 없게 되는 것과 관련된다.In addition, the supplier of fish oil may be unreliable due to the failure or variability of various fisheries. This is related to the inability to use enough fish oil to meet the global demand for DHA.

어유에 대한 대안으로서, PUFA는 미생물로부터 얻을 수 있다. 특히, 해조류는 해양 먹이 사슬에서 오메가-3 다불포화 지방산의 1차 생산자인 것으로 여겨지고 있다. 이들 해양 미생물은 미기재 해양 종의 가장 많은 비율을 나타내며(Colwell 1997), 이들 중, 해양 미세조류가 도코사헥사엔산의 카다란 잠재적인 공급원을 형성하고 있다.As an alternative to fish oil, PUFAs can be obtained from microorganisms. In particular, algae are believed to be the primary producers of omega-3 polyunsaturated fatty acids in the marine food chain. These marine microorganisms represent the largest proportion of unlisted marine species (Colwell 1997), of which marine microalgae form the cardinal potential source of docosahexaenoic acid.

n-3 오일의 잠재적인 공급원은 트라우스토키트리드류라 불리는 미세-종속영양균의 군이다. 이들은 난균류(oomycetes) 및 라비린툴리드류(labyrinthulids)와 함께 스트라메노필라 킹덤(Stramenophila kingdom) 하에 현재 분류된 비광합성(non-photosynthetic), 종속영양 미생물 군이다. 쉬조키트리움(Schizochytrium) 및 트라우스토키트리움(Thraustochytrium) 속(屬)의 구성 종들은 그들의 높은 지질함량 및 높은 레벨의 DHA로 인해 산업상 이용을 위해 잠재적인 오메가-3 생산자로서 연구되어 왔다. 이들 종들은 해양 진균으로서 분류될 경우도 있다. 트라우스토키트리드는 부생균류(saprobe)로서, 혹은 경우에 따라 기생 생물로서 공급되는 미세 종속영양균의 관용명이다. 트라우스토키트리드류는 남극 대륙, 북해, 인도, 일본 및 오스트레일리아로부터 분리된 균주와 함께 넓은 지리학적 분포를 가진다(Reviewed by Lewis et al., 1999). 이들은 살아있는 식물에서 드물게 발견되고 식물 항균제에 의해 억제되는 것으로 보인다. 이들 군의 구성 종은 때로는 거대 조류 및 수생 맨그로브(Mangrove) 잎 등의 죽은 토착성뿐만 아니라 이지성 식물 소재에 풍부하게 존재한다. 이들은 통상 연안 및 심해를 비롯한 원양 수주(water column) 및 침전물에 있다.A potential source of n-3 oils is a group of micro-dependent nutrients called traustokitrides. These fungi I (oomycetes) and La Strada is fishy Drew Tully Menno pillar Kingdom (Stramenophila kingdom) Current classification of non-photosynthetic (non-photosynthetic), under heterotrophic microorganisms with (labyrinthulids). The constituent species of the Schizochytrium and Thraustochytrium genus have been studied as potential omega-3 producers for industrial use due to their high lipid content and high levels of DHA. These species may be classified as marine fungi. Thraustochytrid is the common name for microheterotrophs supplied as a saprobe or, optionally, as a parasitic organism. The traustochytrids have a wide geographic distribution with strains isolated from Antarctica, North Sea, India, Japan and Australia (Reviewed by Lewis et al., 1999). They are rarely found in living plants and appear to be inhibited by plant antimicrobials. The constituent species of these groups are sometimes abundant in the indigenous plant material as well as dead indigenous such as giant birds and aquatic Mangrove leaves. They are usually found in water columns and sediments, including coastal and deep seas.

어유와 비교해서, 미생물은 덜 비린내가 나고 고도로 정제된 DHA 및 기타 PUFA를 가지며 용이하게 양산가능한 지방산의 안정한 공급원을 제공할 수 있다. Compared to fish oil, microorganisms can provide a stable source of fatty acids that are less fishy and have highly purified DHA and other PUFAs.

수개의 최근의 연구는 이하의 1. 내지 3.을 생산하는 트라우스토키트리드 균주의 능력을 목록화하고 있다:Several recent studies have listed the ability of the traustochytrid strain to produce the following 1-3.

1. 배지에서의 고도의 바이오매스1. High Biomass in Medium

2. 이 바이오매스의 일부로서의 고비율의 지질2. High proportion of lipids as part of this biomass

3. 지질에서의 고비율의 PUFA.3. High proportion of PUFA in lipids.

트라우스토키트리드에 의한 PUFA의 생산에 관한 대부분의 보고는 이 화합물이 최근 보고된 많은 트라우스토키트리드 균주에 의해 생산된 가장 풍부한 PUFA이기 때문에 DHA 생산에 대해 거의 배타적으로 다루고 있었다.Most reports on the production of PUFAs by traustochytrid dealt almost exclusively with DHA production since this compound is the most abundant PUFA produced by many of the recently reported traustochytrid strains.

고레벨의 DHA는 예컨대 크립테코디늄 코니(Crypthecodinium cohni) 및 암피디늄(Amphidinium) 종 등의 와편모충류(dinoflagellates)에서도 발견된다. 과학 문헌에 있어서 이용가능한 데이터는 상이한 트라우스토키트리드 균주에 대해 얻어진 바이오매스, 지질 및 최대 DHA 수율에서 편차가 큰 것을 입증한다. 예를 들어, 쉬조키트리움 아그레가툼(Schizochytrium aggregatum)은 10일 후 0.9g/ℓ의 바이오매스를 생산한 반면(Vazhappilly and Chen, 1998), 쉬조키트리움 종 SR21을 이용해서 4일 후에 48g/ℓ의 바이오매스를 얻고 있다(yaguchi et al., 1997). 미생물 PUFA 생산 프로세스의 개발은 적절한 미생물 및 최적화된 배양 기술의 선택을 필요로 하는 것은 매우 명확하다.High levels of DHA, for example, Crypthecodinium It is found in cohni) and cancer PD uranium (Amphidinium) dinoflagellate (dinoflagellates) of such kind. The data available in the scientific literature demonstrate that the deviations in biomass, lipids and maximum DHA yields obtained for different traustochytrid strains are large. For example, Schizochytrium aggregatum produced 0.9 g / l biomass after 10 days (Vazhappilly and Chen, 1998), while 48 g / day after 4 days using Schizochytrium species SR21. l biomass is obtained (yaguchi et al., 1997). It is very clear that the development of the microbial PUFA production process requires the selection of appropriate microorganisms and optimized culture techniques.

PUFA가 풍부한 산물의 증대된 생산을 달성하기 위해서는, 이하의 주된 전략을 타개할 필요가 있다:In order to achieve increased production of PUFA-rich products, it is necessary to break the following main strategies:

1. PUFA-생산 균주의 추가의 분리, 선별 및 유지: DHA-풍부 오일의 상업적 생산을 위한 잠재성을 가진 수개의 균주는 이미 분리되어 있다. 그러나, 보다 높은 PUFA 수율 및/또는 더욱 관심을 끄는 PUFA 프로파일을 생산하는 트라우스토키트리드류가 분리되어 최적화될 수 있다면, 이들 분리주 및 화합물에 대한 수요도 증가할 수 있다.1. Further Isolation, Screening and Maintenance of PUFA-producing Strains: Several strains with potential for commercial production of DHA-rich oils have already been isolated. However, if traustochytrides that produce higher PUFA yields and / or more interesting PUFA profiles can be isolated and optimized, the demand for these isolates and compounds may also increase.

2. PUFA 생산 효율의 최적화: 개별적인 트라우스토키트리드 균주에 의해 생산된 PUFA의 종류와 양은 배양 조건을 변화시킴으로써 조정되기 쉽다. 분자 기술을 이용한 PUFA 프로파일의 증강도 고려될 수 있다. 상이한 시장은 바이오매스(즉, PUFA 생산 w/w 세포 질량) 또는 체적(즉, 생산 w/v 발효 매질)의 관점에서 측정된 높은 레벨의 PUFA를 생산하는 균주에 대한 수요를 제공할 것이다.2. Optimization of PUFA Production Efficiency: The type and amount of PUFAs produced by individual traustochytrid strains are likely to be adjusted by changing the culture conditions. Enhancement of the PUFA profile using molecular techniques can also be considered. Different markets will provide demand for strains that produce high levels of PUFAs measured in terms of biomass (ie, PUFA producing w / w cell mass) or volume (ie, production w / v fermentation medium).

3. 쉬조키트리움 종은 종속영양 배양에 의해 DHA를 생산하는 데 이용된다(OmegaTech, Boulder, Colorado, USA). 쉬조키트리움 균주의 주된 결점은 DHA 이외에도 미생물 오일에 있어서 오메가-6 도코사펜타엔산(DPA)을 생산하는 점이다(Nakhara et al. 1996: Yokochi et al. 1998: Ratledge 2001). DPA의 영양학적 특성은 현재 충분히 공지되어 있지 않으므로, 식품 및 약제학적 용도를 위한 오일 중에서의 그의 존재는 바람직하지 못하다. DHA로부터의 DPA의 분리는 어렵고 비용이 많이 든다.3. Schizochytrium species is used to produce DHA by heterotrophic culture (OmegaTech, Boulder, Colorado, USA). A major drawback of the Schizochytrium strain is the production of omega-6 docosapentaenoic acid (DPA) in microbial oils in addition to DHA (Nakhara et al. 1996: Yokochi et al. 1998: Ratledge 2001). Since the nutritional properties of DPA are not currently well known, their presence in oils for food and pharmaceutical use is undesirable. Separation of DPA from DHA is difficult and expensive.

본 발명은 고아의 역류로부터 회수된 맨그로브라 불리는 해양 관류 식물의 가지 돌기 및 죽은 잎(dead leaves)으로부터 선별된 상당량의 도코사헥사엔산(DHA; C22:6, n-3)을 생산하는 몇몇 균주인 수개의 트라우스토키트리드 균주의 분리에 관한 것이다. 이들 균주는 도코사헥사엔산의 상업적 생산을 위해 주된 중요성을 가질 것이다.The present invention produces a significant amount of docosahexaenoic acid (DHA; C22: 6, n-3) selected from the dendrites and dead leaves of marine perfusion plants called mangroves recovered from the reflux of orphans. Some strains relate to the isolation of several Thraustochytrid strains. These strains will be of major importance for the commercial production of docosahexaenoic acid.

종래 기술:Prior art:

Huang 등은 새롭게 단리된 도코사헥사엔산 생산 트라우스토키트리드류를 그들의 다불포화 지방산 프로파일 및 18S rRNA 유전자의 비교 분석에 의거해서 그룹화하는 것을 기술한 바 있다. 상당량의 DHA를 생산하는 해양 미생물의 7종의 균주를 일본 및 피지의 해안 지역에서 수집한 해수로부터 선별하였다. 이들은 DHA에 부가해서 그들의 중간체인 호흡기 관련 중간체 지방산을 축적하고 있다. 이들 분리주는 18S rRNA 유전자에 있어서의 그들의 특정 삽입 서열에 의해 새로운 트라우스토키트리드인 것으로 입증되었다. 상기 분리주 및 전형적인 트라우스토키트리드류로부터의 18S rRNA 유전자의 분자 분석에 의해 구축된 계통 발생 나무는 동일한 PUFA 프로파일을 가진 균주가 각 단진화 클러스터(monophyletic cluster)를 형성하는 것을 나타낸다. 이들 결과는 C20-22 PUFA 프로파일이 트라우스토키트리드의 그룹화를 위한 유효한 특징으로서 인식될 수 있는 것을 시사한다.Huang et al. Described the grouping of newly isolated docosahexaenoic acid-producing traustochytrides based on their polyunsaturated fatty acid profiles and comparative analysis of 18S rRNA genes. Seven strains of marine microorganisms producing significant amounts of DHA were selected from seawater collected in coastal areas of Japan and Fiji. In addition to DHA, they accumulate their intermediates, the respiratory related intermediate fatty acids. These isolates proved to be new traustokytrids by their specific insertion sequence in the 18S rRNA gene. The phylogenetic tree constructed by molecular analysis of the 18S rRNA genes from the isolates and the typical trastutochytrides shows that strains with the same PUFA profile form each monophyletic cluster. These results suggest that the C20-22 PUFA profile can be recognized as a valid feature for the grouping of traustochytrids.

Fan 등은 문헌[Journal of Industrial Microbiology & Biotechnology (2001)]에 트라우스토키트리드류의 오카라-활용 잠재능(okara-utilizing potential)과 9종의 트라우스토키트리드 균주에 의한 에이코사펜타엔산 및 도코사헥사엔산 생산을 기재한 논문을 발표한 바 있다. 이들 트라우스토키트리드 균주는 아열대 맨그로브로부터 분리된 바 있고, 글루코스 효모 배지 중에서의 상기 지방산의 생산 잠재능을 위해 선별되었다. EPA 및 DHA 생산과 성장을 위해 오카라(두유 잔류물)를 활용하는 그들의 능력도 평가되었다. EPA 수율은 대부분의 균주에서 낮았던 반면, DHA 레벨은 울케니아 종(Ulkenia sp.) KF13을 제외하고 모든 균주에 대해서 총 지방산의 28.1 내지 41.1%를 생산하는 글루코스 효모 추출물 배지 상에서 높았다. 쉬조키트리움 맨그로비에(Schizochytrium mangrovie)의 DHA 수율은 25℃에서 52시간의 발효 후 747.7 내지 2778.9 ㎎/ℓ의 범위였다.Fan et al. Reported in the Journal of Industrial Microbiology & Biotechnology (2001) the eicosapentaenoic acid by the okara-utilizing potential of traustochytrids and nine strains of traustochytrid and A paper describing the production of docosahexaenoic acid was published. These troutstochytrid strains have been isolated from subtropical mangroves and selected for the production potential of these fatty acids in glucose yeast medium. Their ability to utilize okara (soy milk residues) for EPA and DHA production and growth was also evaluated. EPA yield was low in most strains, while DHA levels were high on glucose yeast extract medium producing 28.1 to 41.1% of total fatty acids for all strains except Ulkenia sp. KF13. The DHA yield of Schizochytrium mangrovie ranged from 747.7 to 2778.9 mg / l after 52 hours of fermentation at 25 ° C.

"트라우스토키트리움 종 KK17-3에 의해 생산된 다불포화 지방산의 프로파일"이란 표제 하의 공보는 미끼법에 의해 일본의 이리오모테섬 부근 및 세토 내해의 연안 해수로부터 새롭게 분리된 다불포화 지방산(PUFA)을 생산하는 300종 이상의 미생물 균주의 분리에 관한 것이다. DHA 생산 균주로부터의 PUFA의 프로파일은 4종으로 분류되었다. 다음에, KK17-3이라 불리는 균주는 그의 높은 DHA 함량(총 지방산의 52.1%) 및 DHA뿐만 아니라 아라키돈산, 에이코사펜타엔산 및 도코사펜타엔산을 비롯한 광범위한 기타 PUFA로 인해 추가의 연구를 위해 선정되었다. 18S rRNA 유전자 서열의 분자 계통 발생 분석은 KK17-3이 트라우스토키트리드인 것을 나타내고 있었다.The publication under the heading "Profiles of Polyunsaturated Fatty Acids Produced by Traustochytrium Species KK17-3" uses a decoy method to remove polyunsaturated fatty acids (PUFAs) newly isolated from coastal seawater near Iriomote Island in Japan and the Seto Inland Sea. The isolation of more than 300 microbial strains to produce. The profiles of PUFAs from DHA producing strains were classified into four species. Next, a strain called KK17-3 has further studies due to its high DHA content (52.1% of total fatty acids) and DHA as well as a wide range of other PUFAs including arachidonic acid, eicosapentaenoic acid and docosapentaenoic acid. Was selected for. Molecular phylogenetic analysis of the 18S rRNA gene sequence indicated that KK17-3 was a traustokitride.

Bowles 등에 의한 또 다른 공보는 트라우스토키트리드의 해양 원생생물군의 구성 종에 의한 장쇄 n-3 다불포화 지방산 생산에 관한 것이다. 분리주의 선별을 수행한 후 DHA 생산의 최적화를 수행하였다. 분리 프로그램은 3개의 상이한 지역으로부터 수행되었고, 그들의 바이오매스, 오일 및 도코사헥사엔산 생산(DHA)에 대 해 57개의 분리주를 선별하였다. DHA는 이들 분리주에 존재하는 총 지방산의 50%를 나타내었다. 몇몇 연구는 또한 높은 C:N 비의 배지가 DHA 생산을 자극하는 것을 나타내고 있었다. 최적 DHA 생산은 배양 107시간 후 2.17g/ℓ였다.Another publication by Bowles et al. Relates to the production of long chain n-3 polyunsaturated fatty acids by the constituent species of the marine protozoa of Traustochytrid. Screening of isolates was performed followed by optimization of DHA production. Separation programs were conducted from three different locations and 57 isolates were selected for their biomass, oil and docosahexaenoic acid production (DHA). DHA represented 50% of the total fatty acids present in these isolates. Some studies have also shown that high C: N ratio media stimulate DHA production. Optimal DHA production was 2.17 g / l after 107 hours of culture.

상기 공보는 "해양 미세 조류인 쉬조키트리움 맨그로비에의 지방산 조성 및 스쿠알렌 함량"이란 표제를 달고 있다. 이 공보는 홍콩 맨그로브 서식지에서 부패 중인 칸델리아 칸델(Kandelia candel) 잎으로부터 새롭게 분리된 트라우스토키트리드류인 쉬조키트리움 맨그로비에(S. mangrovie) 중에서의 지방산 조성 및 스쿠알렌 함량의 동정(identification)에 관한 것이다. 3종의 맨그로비에 균주 모두에서의 주된 지방산 구성 성분은 테트라데칸산, 헥사데칸산, 도코사펜탄산 및 도코사헥사엔산이었다. DHA는 가장 주류를 이루고 있는 다불포화 지방산이었고, 이들 균주 모두 중에서의 (총 지방산의) DHA의 비율은 32.29%에서 39.14%까지 다양하였다.The publication is entitled "Facial Fatty Acid Composition and Squalene Content in Marine Microalgae, Schizochytrium mangroves." This bulletin is decaying in the Hong Kong Mangrove Habitat Kandelia. candel ) relates to the identification of fatty acid composition and squalene content in S. mangrovie , the traustochytrids newly isolated from the leaves. The main fatty acid components in all three mangrove strains were tetradecanoic acid, hexadecanoic acid, docosapentanoic acid and docosahexaenoic acid. DHA was the most mainstream polyunsaturated fatty acid, and the proportion of DHA (of total fatty acids) in all of these strains varied from 32.29% to 39.14%.

국제 특허 공개 공보 제WO 98/01536호에는 단기간의 배양 기간 내에 도코사헥사엔산을 생산할 수 있는 쉬와넬라(Shewanella) 속 혹은 슈도알테로모나스(pseudoalteromonas) 속에 속하는 미생물이 기재되어 있다.WO 98/01536 describes microorganisms belonging to the genus Shewanella or pseudoalteromonas , which can produce docosahexaenoic acid in a short period of culture.

본 발명은 고아의 역류로부터 맨그로브라 불리는 해양 관속 식물의 가지돌기 및 죽은 잎에서 살고 있는 10종의 미공표된 트라우스토키트리드 균주의 동정에 관한 것이다. 분리에 이어, 미생물에 대한 GC-MS 분석을 행하여 그들의 총 지방산 함량을 구하였다. 이들 유기체의 18S rRNA 서열은 그들의 계통 발생을 나타낸다. 추가의 연구에 있어서, 이들 유기체는 도코사헥사엔산의 유효한 대량 생산자일 수 있다.The present invention relates to the identification of 10 unpublished traustokytrid strains that live in dead leaves and dendrites of marine tubers called mangroves from the reflux of orphans. Following isolation, GC-MS analysis of the microorganisms was performed to determine their total fatty acid content. The 18S rRNA sequences of these organisms indicate their phylogeny. In further studies, these organisms may be effective mass producers of docosahexaenoic acid.

도 1은 상이한 트라우스토키트리드 균주의 지방산 프로파일을 나타낸 표.1 is a table showing the fatty acid profile of different trastutoquitrid strains.

본 발명의 실시예를 설명하기에 앞서, 참고로 본 명세서에 첨부한 서열목록의 내용은 다음과 같다.Prior to describing the embodiments of the present invention, the contents of the sequence list attached to the present specification are as follows.

서열번호 1은 SC1의 18S rRNA 서열을 나타낸다.SEQ ID NO: 1 shows the 18S rRNA sequence of SC1.

서열번호 2는 AVE1의 18S 리보솜 서열을 나타낸다.SEQ ID NO: 2 shows the 18S ribosomal sequence of AVE1.

서열번호 3은 AVE2의 18S 리보솜 서열을 나타낸다.SEQ ID NO: 3 shows the 18S ribosomal sequence of AVE2.

서열번호 4는 AVE3의 18S 리보솜 서열을 나타낸다.SEQ ID NO: 4 shows the 18S ribosomal sequence of AVE3.

서열번호 5는 AVE4의 18S 리보솜 DNA 서열을 나타낸다.SEQ ID NO: 5 shows the 18S ribosomal DNA sequence of AVE4.

서열번호 6은 AVE5의 18S 리보솜 DNA 서열을 나타낸다.SEQ ID NO: 6 shows the 18S ribosomal DNA sequence of AVE5.

서열번호 7은 AVE6의 18S 리보솜 서열을 나타낸다.SEQ ID NO: 7 shows the 18S ribosomal sequence of AVE6.

서열번호 8은 AVE7의 18S 리보솜 서열을 나타낸다.SEQ ID NO: 8 shows the 18S ribosomal sequence of AVE7.

서열번호 9는 AVE8의 18S 리보솜 DNA 서열을 나타낸다.SEQ ID NO: 9 shows the 18S ribosomal DNA sequence of AVE8.

서열번호 10은 AVE9의 18S 리보솜 서열을 나타낸다.SEQ ID NO: 10 shows the 18S ribosomal sequence of AVE9.

트라우스토키트리움 균주는 맨그로브라 불리는 해양 관상 식물의 가지돌기 및 죽은 잎에서 살고 있다. 이들 식물로부터의 죽은 잎은 고아의 역류로부터 회수되어 폴렌 미끼법에 의해 분리되었다. 상기 잎들은 3일 동안 항생제(스트렙토마이 신 및 페니실린) 및 소나무 꽃가루 알갱이를 구비한 멸균 해수 중에서 배양되었다. 이들 꽃가루는 항생제를 구비한 신선한 멸균 해수 속에 재접종하였다. 접종 3일 후, 꽃가루를 현미경 하에 관찰하였다. 10종의 상이한 트라우스토키트리드 균주는 항생제와 함께 5일 동안 MV(암염(Rock salt)-3.4%, 펩톤-0.15%, 효모 추출물-0.1%, 글루코스-2%, KH₂PO₄-0.025%, pH 7) 한천 플레이트 상에 이식하였다. 상기 트라우스토키트리드류는 상기 플레이트 상에 콜로니/층을 형성하였다. 이들 트라우스토키트리드류는 항생제를 구비한 신선한 MV 배지에서 배양되었다. 성장 3일 후, 순수 배양액이 얻어졌다. 트라우스토키트리드류는 5일의 기간 동안 배양하였고, 세포에 대해서는 총 지방산 함량을 결정하기 위해 GC-MS 분석을 행하였다.Traustochytrium strains live on dendrites and dead leaves of marine ornamental plants called mangroves. Dead leaves from these plants were recovered from the reflux of orphans and separated by the pollen bait method. The leaves were incubated in sterile seawater with antibiotics (streptomycin and penicillin) and pine pollen grains for 3 days. These pollen was reinoculated in fresh sterile seawater with antibiotics. Three days after inoculation, pollen was observed under a microscope. Ten different traustochytrid strains with MV (Rock salt -3.4%, Peptone-0.15%, Yeast Extract -0.1%, Glucose-2%, KH ₂ PO ₄ -0.025% for 5 days with antibiotics) , pH 7) on agar plates. The traustochytrids formed colonies / layers on the plate. These traustokitrides were cultured in fresh MV medium with antibiotics. After 3 days of growth, pure cultures were obtained. Thraustochytrides were cultured for a period of 5 days and cells were subjected to GC-MS analysis to determine total fatty acid content.

많은 양의 Large amount DHADHA 를 생산하는 균주를 동정하기 위한 To identify strains that produce 트라우스토키트리드의Of traustock kitrid 선별. Selection.

총 지질은 2:1(v/v)의 클로로포름:메탄올로 추출하는 폴츠 법(Folch method)을 이용해서 배양액으로부터 추출하였다. 개별 지방산뿐만 아니라 총 지질의 정량화를 위해서, 추출 전에 내부 표준(헵타데칸산 혹은 펜타데칸산)을 첨가하였다. 지방산의 추출 후, 에스터화하고, FID 검출기를 장착한 HP 6850 시리즈 가스 크로마토그래피 상에 아길렌트(Agilent) 5973 질량 분광분석기 및 GC에 접속된 아길렌트 6890 N 가스 크로마토그래피에 의해 GC-MS 분석을 행하였다. 상기 GC-MS 검출은 모세관 칼럼(30 m, HP-5ms, WCOT, 내경: 0.25 ㎜, 막 두께: 0.25 ㎛, 오븐: 150℃에서 2분, 300℃까지 6℃/분, 300℃에서 20분, 헬륨 캐리어 가스 유량: 1.0 ㎖/ 분, 분리비 50:1)을 가진 EI 모드에 있어서 70 eV(m/z 50-550; 230℃에서 소스 및 150℃에서 사중극)에서 수행하였다. GC-FID를 위해서, 모세관 칼럼 DB-23(30m, WCOT, 내경: 0.25 mm, 막 두께: 0.5 ㎛)을 이용하였다. 오븐 온도는 160℃에서 2분, 180℃까지 6℃/분, 180℃에서 2분, 230℃까지 4℃/분 및 230℃에서 10분으로 프로그래밍되어 있었고, 이때, N₂ 캐리어 가스 유량은 1.5 ㎖/분, 주입기 온도는 230℃, 검출기 온도는 250℃, 분리비는 50:1이었다.Total lipids were extracted from the cultures using the Folch method extracted with 2: 1 (v / v) chloroform: methanol. For quantification of individual lipids as well as total lipids, internal standards (heptadecanoic acid or pentadecanoic acid) were added before extraction. After extraction of fatty acids, GC-MS analysis was carried out by agilent 5973 mass spectrometer and GC-connected Agilent 6890 N gas chromatography on HP 6850 series gas chromatography equipped with FID detector. It was done. The GC-MS detection was performed using a capillary column (30 m, HP-5 ms, WCOT, inner diameter: 0.25 mm, film thickness: 0.25 μm, oven: 150 ° C., 2 minutes, 300 ° C., 6 ° C./minute, 300 ° C., 20 minutes). And 70 eV (m / z 50-550; source at 230 ° C. and quadrupole at 150 ° C.) in EI mode with helium carrier gas flow rate: 1.0 mL / min, separation ratio 50: 1. For GC-FID, capillary column DB-23 (30 m, WCOT, inner diameter: 0.25 mm, membrane thickness: 0.5 μm) was used. The oven temperature was programmed to 2 minutes at 160 ° C., 6 ° C./minute up to 180 ° C., 2 minutes at 180 ° C., 4 ° C./minute up to 230 ° C. and 10 minutes at 230 ° C., with a N ₂ carrier gas flow rate of 1.5 Ml / min, injector temperature was 230 ° C., detector temperature was 250 ° C., and separation ratio was 50: 1.

분리된 10개의 균주의 지방산 프로파일은 도 1에 표시되어 있다.Fatty acid profiles of the 10 isolated strains are shown in FIG. 1.

이들 균주의 지방산 프로파일은 AVE5, AVE7 및 SC-1이 상당한 양의 DHA를 생산하는 것을 시사한다. AVE1은 또한 DHA의 유망한 생산자이다. 그러나, 기타 균주는 다량의 DHA를 생산하지 못한다.The fatty acid profile of these strains suggests that AVE5, AVE7 and SC-1 produce significant amounts of DHA. AVE1 is also a promising producer of DHA. However, other strains do not produce large amounts of DHA.

트라우스토키트리드의Of traustock kitrid 18s  18s rRNArRNA 서열분석 Sequencing

높은 DHA 함량을 가진 SC-1, AVE5 및 AVE7의 1.7kb 18s rRNA 서열은 프라이머를 이용해서 게놈으로부터 보존 영역까지 증폭시켰다. 증폭된 단편을 서열 분석하였다. 이 서열은 서열목록에 표시되어 있다. 따라서, 3종의 균주, 즉, SC-1, AVE5 및 AVE7은 DHA의 생산자인 것으로 보인다. 이들 균주는 인도의 찬디가르에 소재한 IMTECH(Institute of Microbial Technology)의 MTCC(Microbial Type Culture Collection and Gene Bank)에 기탁되어 있다.The 1.7 kb 18s rRNA sequences of SC-1, AVE5 and AVE7 with high DHA content were amplified from the genome to the conserved region using primers. Amplified fragments were sequenced. This sequence is shown in the sequence listing. Thus, three strains, SC-1, AVE5 and AVE7, appear to be producers of DHA. These strains have been deposited in the Microbial Type Culture Collection and Gene Bank (MTCC) of the Institute of Microbial Technology (IMTECH) in Chandigarh, India.

추가의 연구에서의 이들 유기체는 DHA의 다량의 상업적 생산에 있어 매우 효능이 있는 것임을 입증할 수 있다.These organisms in further studies may prove to be very potent in the commercial production of large amounts of DHA.

<110> AVESTHAGEN LIMITED <120> DOCOSAHEXAENOIC ACID (DHA) PRODUCING THRAUSTOCHYTRID STRAIN-SC1 <160> 10 <170> KopatentIn 1.71 <210> 1 <211> 1547 <212> DNA <213> Thraustochtrid <220> <221> rRNA <222> (1)..(1547) <400> 1 tacctggttg atcctgccag tagtcatatg ctcgtctcaa agattaagcc atgcatgtgt 60 aagtataagc gattgtactg tgagactgcg aacggctcat tatatcagta ataatttctt 120 cggtagtttc ttttatatgg atacctgcag taattctgga aataatacat gctgtaagag 180 ccctgtatgg ggctgcactt attagattga agccgatttt attggtgagt catgataatt 240 gagcagattg actattttgg tcgatgaatc gtttgagttt ctgccccatc agttgtcgac 300 ggtagtgtat tggactacgg tgactataac gggtgacgga gagttagggc tcgactccgg 360 agagggagcc tgagagacgg ctaccatatc caaggatagc agcaggcgcg taaattaccc 420 actgtggact ccacgaggta gtgacgagaa atatcgatgc gaagcgtgta tgcgttttgc 480 tatcggaatg agagcaatgt aaaaccctca tcgaggatca actggagggc aagtctggtg 540 ccagcagccg cggtaattcc agctccagaa gcatatgcta aagttgttgc agttaaaaag 600 ctcgtagttg aatttctggc atgggcgacc ggtgctttcc ctgaatgggg attgattgtc 660 tgtgttgcct tggccatctt tttcttttct ttttagggga gaaatctttc actgtaatca 720 aagcagagtg ttccaagcag gtcgtatgac cggtatgttt attatgggat gataagatag 780 gacttgggtg ctattttgtt ggtttgcacg cctgagtaat ggttaatagg aacagttggg 840 ggtattcgta tttaggagct agaggtgaaa ttcttggatt tccgaaagac gaactagagc 900 gaaggcattt accaagcatg ttttcattaa tcaagaacga aagtctgggg atcgaagatg 960 attagatacc atcgtagtct agaccgtaaa cgatgccgac ttgcgattgt tgggtgcttt 1020 tttaatgggc ctcagcagca gcacatgaga aatcaaagtc tttgggttcc ggggggagta 1080 tggtcgcaag gctgaaactt aaaggaattg acggaagggc accaccagga gtggagcctg 1140 cggcttaatt tgactcaaca cgggaaaact taccaggtcc agacataggt aggattgaca 1200 gattgagagc tctttcatga ttctatgggt ggtggtgcat ggccgttctt agttggtgga 1260 gtgatttgtc tggttaattc cgttaacgaa cgagacctcg gcctactaaa tagtgcgtgg 1320 tatggcaaac atagtacgtt tttaacttct tagagggaca tgtccggttt acgggcagga 1380 aagttcgagg caataacagg tctgtgatgc ccttagatgt tctgggccgc acgcgcgcta 1440 cactgatggg ttcatcgggt tttaatttca atttttggaa ttgagtgctt ggtcggaagg 1500 cctggctaat ccttggaacg ctcatcgtgc tggggctaga ttttgca 1547 <210> 2 <211> 1840 <212> DNA <213> Thraustochtrid <400> 2 gctccacgcg ttgggagctc tcccctatgg tcgacctgca ggcggccgcg aattcactag 60 tgatttacct ggttgatcct gccagtagcc ctacgctcgt ctcaaagact aagccatgca 120 tgtgtaagta taagcgaatt atactgtgaa actgcgaacg gctcattata tcagttataa 180 tcccttcggt agttccttta cacggatacc tgcagtaatt ctggaattaa tacgtgctgt 240 acgggcccga ctttcgggga gggccgcact tattaggtct aagccaacgt tattggtgag 300 tcatgataat tgagcagatc gcttttcgga gcgatgaatc gtttgagttt ctgccccatc 360 agttgtcgac ggtagggtat tggcctacgg tgactataac gggtgacggg gagttagggc 420 tcgactccgg agagggagcc tgagagacgg ctaccacatc caaggaaggc agcaggcgcg 480 taaattaccc aatgtggact ccacgaggta gtgacgagaa atatcaatgc ggggcgcttc 540 gcgtcttgct attggaatga gagcaatgta aaaccctcat cgaggatcaa ctggagggca 600 agtctggtgc cagcagccgc ggtaattcca gctccagaag cgtatgctaa agttgttgca 660 gttaaaaagc tcgtagttga atttctggtg tgggagccca ggcctcggtg cgaatgcgcc 720 ttgtactgct ttgcggctcc tttgccatcc tcgtttttcg taagaaaggc gtcattcact 780 gtaatcaaag cagagtgttc caagcaggcc gtagggccgg tatgtttatt atgggatgat 840 cagataggac tcgggtgcta ttttgttggt ttgcacatct gagtaatgat taataggaac 900 agtcgggggt atccgtattt aggagctaga ggtgaaattc ttggatttcc gaaagacgaa 960 ctacagcgaa ggcattacca agcatgtttt cattaatcaa gaacgaaagt ctggggatcg 1020 aagatgatta gataccatcg tagtctagac cgtaaacgat gccgacttgc gattgcgggt 1080 ggcttgtatt gggcctccgc agcagcacat gagaaatcaa agtctttggg ttccgggggg 1140 agtatggtcg caaggctgaa acttgaagga attgacggaa gggcaccacc aggagtggag 1200 cctgcggctt aatttgactc aacacgggaa aacttaccag gtccagacat agtaggattg 1260 acagattgag agctctttct tgattctatg ggtggtggtg catggccgtt cttagttggt 1320 ggagtgattt gtctggttaa ttccgttaac gaacgagacc tcggcctact aaatagccgg 1380 gcgtatggcg acatatgtgt ttgtggcttc ttagagggac atgttcggtt tacgagcagg 1440 aagttcgagg caataacagg tctgtgatgc ccttagatgt tctgggccgc acgcgcgcta 1500 cactgatggg ttcagcgggt cttgttgtga tttatcgcag cgttgctttg tcggaaggca 1560 tggctaatcc tttgaacgcc catcgtgctg gggctagatt tttgcaatta ttaatctcca 1620 acgaggaatt cctagtaaac gcaagtcatc agcttgcatt gaatacgtcc ctgccctttg 1680 tacacaccgc ccgtcgcacc taccgattga acggtccgat gaaaccatgg gactaccttt 1740 tgagcgtttg ttcgcgatgg aggtgggaac tcgggtgaat cttattgttt agaggaaggt 1800 gaagtcgtaa caaggtttcc gtaggtgaac ctgcggaagg 1840 <210> 3 <211> 1798 <212> DNA <213> Thraustochytrid <400> 3 tacctggttg atcctgccag taatcatacg cttgtctcaa agattaagcc atgcatgtct 60 aagtataaag gattatactc tgaaactgcg aacggctcat taaaccagta ctaacctaat 120 tgatgatgaa agttagatgg atacttgtgg caaatctaga accaatacat gcttgggggc 180 ccgactttat gggagggctg catttatttg aaccaaacca atagctcgtt tcggcgggtt 240 tcgtgtgggt gagtcagaat aagtaagcga accctttttt cggagagggt gaatcattcg 300 agtttctgcc ccatcagttg tcgacgacac ggtattgacc tgtcgtgact gtcacgggtg 360 acggagaatt agggttcgat tccggagagg gagcctgaga gacggctacc acatccaagg 420 aaggcagcag gcgcgtaaat tactcaatgt taaatggacg aagtagtgac gagaaataac 480 aatgtggagc gctttgcgtt ttacaattgt aatgagagca gattaaaggg gtcatcgagg 540 atccattgga gggcaagtct ggtgccagca gccgcggtaa ttccagctcc aatagcgtat 600 attaaagttg ttgcagttaa aaagctcgta gttgaagaag ggttgtcttt ggtttggttt 660 tcgtatgaga actgaacatg gcaacattcg tctgtttttg aaagcgggca gaccgtttac 720 tgtaaaaaaa ttagagtgtt caaagcgtga tgtatgttgt ttgaatatag tagtatggaa 780 taatgagata ggactttgtt gttattttgt tggtttgcat ggcaaggtaa tgattaatag 840 ggacagttgg gggtattcgt attacgatgt cagaggtgaa attcttggat tttcgaaaga 900 cgaacaactg cgaaagcatt taccaaggat gttttcatta atcaagaacg aaagtcaggg 960 gatcgaagat gattagatac catcgtagtc ttgaccgtaa acaatgccga cttgcgattg 1020 tgttcttctt gttttagaga gggatgcagc agcacatgag aaatcaaagt ctttgggttc 1080 cggggggagt atggtcgcaa ggctgaaact taaaggaatt gacggaaggg caccaccagg 1140 agtggagcct gcggcttaat ttgactcaac acggggaaac ttaccaggtc cggacatagg 1200 aaggattgac agattgagag ctctttcttg attctatggg tggtggtgca tggccgttct 1260 tagttggtgg agtgatttgt ctggttaatt ccgttaacga acgagaccac agcctactaa 1320 ataatggtgt ttttttgttt taggaaagac atgtcgtttc ttagagggac atttcgggtt 1380 taccggaagg gagtttgtgg caataacagg tctgtgatgc ccttagatgt tctgggccgc 1440 acgcgcgcta cactgatcgg ttcaacgagt gtttgcagtt tcgttttgag actgtgttcc 1500 ttggtaggaa tgcctgggta atcttttgaa cgccgatcgt gatggggcta gatttttgca 1560 attgttaatc tccaacgagg aattcctagt aaacgcaagt catcagcttg cattgattac 1620 gtccctgccc cttgtacaca ccgcccgtcg cacctaccga ttgaacgatc cggtgagacc 1680 ttgggattgc tcgattttct tttcacgagg agagtcagta gcaagaactt gagcaaacct 1740 tatcgtttag aggaaggtga agtcgtaaca aggtttccgt aggtgaacct gcggaagg 1798 <210> 4 <211> 1881 <212> DNA <213> Thraustochytrid <400> 4 acgcgttggg agctctccca tctggtcgac ctgcaggcgg ccgcgaattc actagtgatt 60 ccttccgcag gttcacctac ggaaaccttg ttacgacttc accttcctct aaacgataag 120 gtttactcaa cttttcccgt catcacacga acaaacgcaa aacaacagga tccgaagatt 180 tcaccggatc gttcaatcgg taggtgcgac gggcggtgtg tacaaagggc agggacgtaa 240 tcaatgcaag ctgatgactt gcgtttacta ggaattcctc gttggagatt agaattgcaa 300 aaatctagcc ccatcacgat ggacatttaa aagattgccc ggacattcct gtctaggacc 360 gtcattcatc gttaaataaa caacacatac tcgttgaatc catcagtgta gcgcgcgtgc 420 ggcccagaac atctaagggc atcacagacc tgttattgcc acaaacttcc ttccggtaaa 480 accgaaatgt ccctctaaga agtctcgcca ctatgtcacc atagcacgta ctatttagta 540 ggctgtggtc tcgttcgtta acggaattaa ccagacaaat cactccacca actaagaacg 600 gccatgcacc accacccata gaatcaagaa agagctctca atctgtcaat ccttcctatg 660 tctggacctg gtaagttttc ccgtgttgag tcaaattaag ccgcaggctc cactcctggt 720 ggtgcccttc cgtcaattcc tttaagtttc agccttgcga ccatactccc cccggaaccc 780 aaagactttg atttctcatg tgctgctgcc caagcctatt tcaaaaaaac atcggacaat 840 cgcaagtcgg catcgtttac ggttaagact acgatggtat ctaatcatct tcgatcccct 900 aactttcgtt cttgattaat gaaaacatcc ttggtaaatg cttttcgcag ttgttcgtct 960 tttgtaaatc caagaatttc acctctgaca accaaatacg aataccccca actgtccctg 1020 ttaatcatta ctttgatgtg caaaccaaca aaatagcaac aaagtcctat cttattattc 1080 catactactc tattccaacc atacgatttg cttgaaacac tctaattttt ttacagtaaa 1140 tgatgcccac ccacgcccaa aggcgcaaac gagcaaccat ggccagatca aacacaaaag 1200 ccactctcag cattcgccga aaagcctggt ccgtcctacc aatacttcaa ctacgagctt 1260 tttaactgca acaactttaa tacacgctac cggagctgga gttaccgcgg ctgctggcac 1320 cagacttgcc ctccagttga tcctcgatga gggttttaga ttgctctcat tacagaaaga 1380 gtaggcttag cccaccccac tgttatttct cgtcactact tcgtcgaatt gacattgagt 1440 aatttgcgcg cctgctgcct tccttggatg gggtagccgt ctctcaggct ccctctccgg 1500 aatcgaaccc tgattctccg tcacccgtaa caatcatggt aggccaatac cctaccatcg 1560 acaattgatg gggcagaaac ttgaatgaac atcgccggca caaacacctc tgcgattcgc 1620 ctagttacta tgaatcacca aaacaacccc gagaggtatt ggtttatatc taataaatgc 1680 agcccttccc aaaagtcggg ccctaaagca tgtattattt ctagatttgc cacaagtatc 1740 catctaatca aacattttca aagaaactat aactgtttta atgagccgtt cgcagtttca 1800 ccgtataaat cgcttatact tagacatgca tggcttagtc tttgagacaa gcgtgcgatt 1860 actggcagga tcaaccaggt a 1881 <210> 5 <211> 1795 <212> DNA <213> Thraustochytrids <400> 5 ccttccgcag gttcacctac ggaaaccttg ttacgacttc accttcctct aaacaataag 60 attcacccga gttctgcctc tgtccaaaaa tcaatccaaa cagaaacatc ccatggtttc 120 atcggaccgt tcaatcggta ggtgcgacgg gcggtgtgta caaagggcag ggacgtattc 180 aatgcaagct gatgacttgc gtttactagg aattcctcgt tggagattaa taattgcaaa 240 aatctagccc cagcacgatg agcgttccaa ggattagcca ggccttccga ccaagcactc 300 aattccaaaa attagaatta aaacccgatg aacccatcag tgtagcgcgc gtgcggccca 360 gaacatctaa gggcatcaca gacctgttat tgcctcgaac ttcctgcccg taaaccggac 420 atgtccctct aagaagtaaa aacgcactat gttgccatac cacgcactat ttagtaggcc 480 gaggtctcgt tcgttaacgg aattaaccag acaaatcact ccaccaacta agaacggcca 540 tgcaccacca cccatagaat catgaaagag ctctcaatct gtcaatccta cctatgtctg 600 gacctggtaa gttttcccgt gttgagtcaa attaagccgc aggctccact cctggtggtg 660 cccttccgtc aattccttta agtttcagcc ttgcgaccat actccccccg gaacccaaag 720 actttgattt ctcatgtgct gctgctgagg cccatataat aaagcaccca acaatcgcaa 780 gtcggcatcg tttacggtct agactacgat ggtatctaat catcttcgat ccccagactt 840 tcgttcttga ttaatgaaaa catgcttggt aaatgccttc gctctagttc gtctttcgga 900 aatccaagaa tttcacctct agctcctaaa tacgaatacc cccaactgtt cctattaacc 960 attactcagg cgtgcaaacc aacaaaatag cacccaagtc ctatcttatc atcccataat 1020 aaacataccg gtcatacgac ctgcttggaa cactctgctt tgattacagt gaaagatctc 1080 ataccaaaat agcatgagaa agatggccaa ggcaacacag acaatcaatc cccattcagg 1140 gaaagcaccg gtcgcccatg ccagaaattc aactacgagc tttttaactg caacaacttt 1200 agcatatgct tctggagctg gaattaccgc ggctgctggc accagacttg ccctccagtt 1260 gatcctcgat gagggtttta cattgctctc attccgatag caaaacgcat acacgcttcg 1320 catcgatatt tctcgtcact acctcgtggg gtccacagtg ggtaatttac gcgcctgctg 1380 ctatccttgg atatggtagc cgtctctcag gctccctctc cggagtcgag ccctaactct 1440 ccgtcacccg ttatagtcac cgtagtccaa tacactaccg tcgacaactg atggggcaga 1500 aactcaaacg attcatcgac aaaaaagtca atctgctcaa ttatcatgat tcaccaataa 1560 aatcggcttc aatctaataa gtgcagcccc atacagggct cttacagcat gtattatttc 1620 cagaattact gcaggtatcc atataaaaga aactaccgaa gaaattatta ctgatataat 1680 gagccgttcg cagtctcaca gtacaatcgc ttatacttac acatgcatgg cttaatcttt 1740 gagacgagca tatgactact ggcaggatca accaggtaaa tcgaattccc gcggc 1795 <210> 6 <211> 1779 <212> DNA <213> Thraustochytrid <220> <221> rRNA <222> (1)..(1779) <400> 6 tacctggttg atcctgccag tagtcatatg ctcgtctcaa agattaagcc atgcatgtgt 60 aagtataagc gattgtactg tgagactgcg aacggctcat tatatcagta ataatttctt 120 cggtagtttc ttttatatgg atacctgcag taattctgga aataatacat gctgtaagag 180 ccctgtatgg ggctgcactt attagattga agccgatttt attggtgaat catgataatt 240 gagcagattg acttttttgt cgatgaatcg tttgagtttc tgccccatca gttgtcgacg 300 gtagtgtatt ggactacggt gactataacg ggtgacggag agttagggct cgactccgga 360 gagggagcct gagagacggc taccatatcc aaggatagca gcaggcgcgt aaattaccca 420 ctgtggaccc cacgaggtag tgacgagaaa tatcgatgcg aagcgtgtat gcgttttgct 480 atcggaatga gagcaatgta aaaccctcat cgaggatcaa ctggagggca agtctggtgc 540 cagcagccgc ggtaattcca gctccagaag catatgctaa agttgttgca gttaaaaagc 600 tcgtagttga atttctggca tgggcgaccg gtgctttccc tgaatgggga ttgattgtct 660 gtgttgcctt ggccatcttt ctcatgctat tttggtatga gatctttcac tgtaatcaaa 720 gcagagtgtt ccaagcaggt cgtatgaccg gtatgtttat tatgggatga taagatagga 780 cttgggtgct attttgttgg tttgcacgcc tgaataatgg ttaataggaa cagttggggg 840 tattcgtatt taggagctag aggtgaaatt cttggatttc cgaaagacga actagagcga 900 aggcatttac ccaagcatgt tttcattaat caagaacgaa agtctgggga tcgaagatga 960 ttagatacca tcgtagtcta gaccgtaaac gatgccgact tgcgattgtt gggtgcttta 1020 ttatatgggc ctcagcagca gcacatgaga aatcaaagtc tttgggttcc ggggggagta 1080 tggtcgcaag gctgaaactt aaaggaattg acggaagggc accaccagga gtggagcctg 1140 cggcttaatt tgactcaaca cgggaaaact taccaggtcc agacataggt aggattgaca 1200 gattgagagc tctttcatga ttctatgggt ggtggtgcat ggccgttctt agttggtgga 1260 gtgatttgtc tggttaattc cgttaacgaa cgagacctcg gcctactaaa tagtgcgtgg 1320 tatggcaaca tagtgcgttt ttacttctta gagggacatg tccggtttac gggcaggaag 1380 ttcgaggcaa taacaggtct gtgatgccct tagatgttct gggccgcacg cgcgctacac 1440 tgatgggttc atcgggtttt aattctaatt tttggaattg agtgcttggt cggaaggcct 1500 ggctaatcct tggaacgctc atcgtgctgg ggctagattt ttgcaattat taatctccaa 1560 cgaggaattc ctagtaaacg caagtcatca gcttgcattg aatacgtccc tgccctttgt 1620 acacaccgcc cgtcgcacct accgattgaa cggtccgatg aaaccatggg atgtttctgt 1680 ttggattgat ttttggacag aggcagaact cgggtgaatc ttattgttta gaggaaggtg 1740 aagtcgtaac aaggtttccg taggtgaacc tgcggaagg 1779 <210> 7 <211> 1803 <212> DNA <213> Thraustochytrid <220> <221> rRNA <222> (1)..(1803) <400> 7 ccttccgcag gttcacctac ggaaaccttg ttacgacttc accttcctct aaacgttaag 60 gtttgctcaa ctttggccac gaactcctgt cgccaggaac aacgtgccct ccgaaggtct 120 caccggaacg ttcaatcggt aggtgcgacg ggcggtgtgt acaaagggca gggacgtaat 180 caatgcaagc tgatgacttg cgtttactag gaattcctcg ttggagatca ataattgcaa 240 agatctagcc ccatcacgac tggtattcac acgattaccc aaaccttccg gcccaggact 300 ctgtcataac aacaaaaaca acgtgctccc ccagtcagtg tagcgcgcgt gcggcccaga 360 acatctaagg gcatcacaga cctgttattg ccacaaactt ccctctggta aacccaaagt 420 gtccctctaa gaagtgccat accactgtgt cgccacaacg tgcactattt agtaggctgt 480 ggtctcgttc gttaacggaa tcaaccagac aaatcactcc accaactaag aacggccatg 540 caccaccacc catagaatca agaaagagct ctcaatctgt caatccttcc tatgtctgga 600 cctggtaagt ttccccgtgt tgagtcaaat taagccgcag gctccactcc tggtggtgcc 660 cttccgtcaa ttcctttaag tttcagcctt gcgaccatac tccccccgga acccaaagac 720 tttgatttct catgcgctgc cgcctcccgc ttcaagagcc gggagacaat cgcgagtcgg 780 catcgtttac ggtatggact acgatggtat ctaatcatct tcgatccccc atactttcgt 840 tcttgatcaa tgaaaacatc cttggtaaat gctttcgcag tcgttcggcc ttctagtaat 900 ccaagaattt cacctctgac actagaattc gaataccccc aactgtccct cttcatcatt 960 ccgccaacgc gcaaaccaac aaaataccgc caacggccta tctcatcatc cccatactat 1020 catattccaa tatcaacctt tgcttgaaac accctaattt ttttacagtg aaagaggcaa 1080 gtcttacaac ttgccagact ggcagcgcac gtgtcactgt tcggcaagca ttcacccgcc 1140 gacggacacg cacacaaacc atacgttcga ctacgagctt tttaactgca acaactttag 1200 tatacgctat tggagctgga attaccgcgg ctgctggcac cagacttgcc ctccaattga 1260 tccccgatgg gggttttaaa tcgctctcat ttcagtcgag atacgcacag cgcacctcac 1320 tgctatttct cgtcactact tcgttctgta aacttgcagt aatttacgcg cctgctgcct 1380 tccttggatg tggtagccgt ctctcaggct ccctctccgg actcgaaccc taattccccg 1440 tcacccgtaa cggtcacggg aggccaatac cctcccgtcg acaactgatg gggctgaaac 1500 ttgaatgatt catcgccgcc cgagggctat gcgattcgct caattactat gactcaccat 1560 cacaaggccc gagagcctac tggtttaggt ctaataagtg cggccctccc caaaagtcgg 1620 gcctccaagc acgtattgat tctagatttg ccacatgtat ccgtctacac aacgcacaat 1680 catgggaatt ataacagata taatgagccg ttcgcggttt cgctgtatta ccaactaata 1740 ctatcgcatg catggcttaa tctttaagac aagcgtatga ccgctggcag gatcaaccag 1800 gta 1803 <210> 8 <211> 1779 <212> DNA <213> Thraustochytrid <220> <221> rRNA <222> (1)..(1779) <400> 8 tacctggttg atcctgccag tagtcatatg ctcgtctcaa agattaagcc atgcatgtgt 60 aagtataagc gattgtactg tgagactgcg aacggctcat tatatcagta ataatttctt 120 cggtagtttc ttttatatgg atacctgcag taattctgga aataatacat gctgtaagag 180 ccctgtatgg ggctgcactt attagattga agccgatttt attggtgaat catgataatt 240 gagcagattg acttttttgg tcgatgaatc gtttgagttt ctgccccatc agttgtcgac 300 ggtagtgtat tggactacgg tgactataac gggtgacgga gagttagggc tcgactccgg 360 agagggagcc tgagagacgg ctaccatatc caaggatagc agcaggcgcg taaattaccc 420 actgtggact ccacgaggta gtgacgagaa atatcgatgc gaagcgtgta tgcgttttgc 480 tatcggaatg agagcaatgt aaaaccctca tcgaggatca actggagggc aagtctggtg 540 ccagcagccg cggtaattcc agctccagaa gcatatgcta aagttgttgc agttaaaaag 600 ctcgtagttg aatttctggc atgggcgacc ggtgctttcc ctgaatgggg attgattgtc 660 tgtgttgcct tggccatctt tttcttttct ttttagggga gaaatctttc actgtaatca 720 aagcagagtg ttccaagcag gtcgtatgac cggtatgttt attatgggat gataagatag 780 gacttgggtg ctattttgtt ggtttgcacg cctgagtaat ggttaatagg aacagttggg 840 ggtattcgta tttaggagct agaggtgaaa ttcttggatt tccgaaagac gaactagagc 900 gaaggcattt accaagcatg ttttcattaa tcaagaacga aagtctgggg atcgaagatg 960 attagatacc atcgtagtct agaccgtaaa cgatgccgac ttgcgattgt tgggtgcttt 1020 tttatgggcc tcagcagcag cacatgagaa atcaaagtct ttgggttccg gggggagtat 1080 ggtcgcaagg ctgaaactta aaggaattga cggaagggca ccaccaggag tggagcctgc 1140 ggcttaattt gactcaacac gggaaaactt accaggtcca gacataggta ggattgacag 1200 attgagagct ctttcatgat actatgggtg gtggtgcatg gccgttctta gttggtggag 1260 tgatttgtct ggttaattcc gttaacgaac gagacctcgg cctactaaat agtgcgtggt 1320 atggcaacat agtacgtttt tacttcttag agggacatgt ccggtttacg ggcaggaagt 1380 tcgaggcaat aacaggtctg tgatgccctt agatgttctg ggccgcacgc gcgctacact 1440 gatgggttca tcgggtttta attctgtttt tatggaattg agtgcttggt cggaaggcct 1500 ggctaatcct tggaacgctc atcgtgctgg ggctagattt ttgcaattat taatctccaa 1560 cgaggaattc ctagtaaacg caagtcatca gcttgcattg aatacgtccc tgccctttgt 1620 acacaccgcc cgtcgcacct accgattgaa cggtccgatg aaaccatggg atgtttctgt 1680 ttggattgat ttttggacag aggcagaact cgggtgaatc ttattgttta gaggaaggtg 1740 aagtcgtaac aaggtttccg taggtgaacc tgcggaagg 1779 <210> 9 <211> 1793 <212> DNA <213> Thraustochytrid <400> 9 ccttccgcag gttcacctac gaaaaccttg ttacgacttc tccttcctct atgtgctgtg 60 gttcaccaaa ctttccactc ctcaaagaga gactcttatg aggaacggtc cgaattattc 120 accgaagcac acaatcggta ggagcgacgg gcggtgtgta caaagggcag ggacgtaatc 180 aatacgagat gttgactcgt gtttactagg aattcctcgt tgaagactaa taattgcaag 240 agtctatccc catcacgatg cattttaaag agattaccca acccttccag gcaaggaaat 300 acactcgttg aatgcatcag tgtagcgcgc gtgcagccca gaacatctaa gggcatcaca 360 gacctgttat tgcctcagac ttccttgcat tgaacataca aagtccctct aagaagctta 420 cccacatagc ctaggccatg tgtagctatt tagtaggctg aggtctcgtt cgttaacgga 480 attaaccaga caaatcactc caccaactaa gaacggccat gcaccaccac ccatagaatc 540 aggaaagagc tatcaatctg tcaatccttc caatgtctgg atctggtaag tttccccgtg 600 ttgagtcaaa ttaagccgca ggctccactc ctggtggtgc ccttccgtca attcctttta 660 agtttcagcc ttgcgaccat actcccccca gaacccaaag acttggattt ctcatcaagg 720 tgctgagaga gcaataatga cactcacccc aatctctagt tggcatggtt catggctaag 780 actaggacgg tatctaatcg tcttcgatcc cctaactttc cgttcttgat caaagaaaac 840 atccttggca aatgctttcg cattagtgtg tctttaacaa atccaagaat ttcacctctg 900 acagttaaat acaaatgccc ccaattgtcc ctatcaatcg ttgcgatggt tctcgaaacc 960 aataaaaaag atccaaagct caatttcatt attccatgct aatatattca aggcacatgc 1020 ctgcttgaaa cactctaatt tcctcaaagt aaaatagctg acaactatac gaccagcgaa 1080 ggccctatag tcatcaagac tcatgcacag aacatgtcag atactcaccc aggggatgga 1140 ccaacagcct ctatgcagaa attcaactac gagcttttta actgcaacaa ctttaatata 1200 cgctattgga gctggaatta ccgcggctgc tggcaccaga cttgccctcc aattgatact 1260 tgcaaagtgt tttaaattct gctcattcca attacaagac atggatgccc tgtattgtta 1320 tttcttgtca ctacctcctt gtgtcaagat tgggtaattt gcgcgcctgc tgccttcctt 1380 agatgtggta gccatttctc aggctccctc tccggaatcg aaccctaatt ctccgttacc 1440 cgtcattgcc atggtaggcc aataccctac catccaaagc tgataggtca gaaactcatg 1500 agatgcatca ccagaaagac catgtgattc atcctcttac catacttcat tatgatacca 1560 agcaagcttg gattggttct gaatttggta agcaccaccc tccaataagt tgggtgttga 1620 aacatgtatt ggctccagaa ttactgtagt tatccatata acgataatca tcaaatagat 1680 cattactgtt ataatgagcc attcgcagct taaccgtata aaggcataga ctcagacatg 1740 catggcttaa tctttgagac aagcatatga ctactggcag gatcaaccag gta 1793 <210> 10 <211> 1754 <212> DNA <213> thraustochytrid <400> 10 tccttccgca ggttcaccta cggaaacctt gttacgactt caccttcctc taaacgataa 60 gattcgtcga agttcaccgc gcgagccaag tcgccccaac tcgcgcagct cccgacgcct 120 catcggaccg ttcaatcggt aggtgcgacg ggcggtgtgt acaaagggca gggacgtatt 180 cgatgcgagc tgatgactcg cgtctactag gaattcctcg ttggagacca ataattgcaa 240 aggtctagcc ccagcacgac gagcgttcac aggattcgcc gccccctccg aggaagctca 300 gctccgaaaa gcaacacccg atgaacccgt cagtgtagcg cgcgtgcggc ccagaacatc 360 taagggcatc acagacctgt tattgcctcg aacttcccgc tcgtataccg aacatgtccc 420 tctaagaagc ctgcaccatg tcgccatgcc gcagcctagt tagtaggccg aggtctcgtt 480 cgttaacgga attaaccaga caaatcactc caccaactaa gaacggccat gcaccaccac 540 ccatagaatc atgaaagagc tctcaatctg tcaatcctac ctatgtctgg acctggtaag 600 tttccccgtg ttgagtcaaa ttaagccgca ggctccactc ctggtggtgc ccttccgtca 660 attcctttaa gtttcagcct tgcgaccata ctccccccgg aacccaaaga ctctggtttc 720 ccacacgctc ccgcagaagc tcacaagagc accctgcgat cgcgggtcgg catcgtttgc 780 ggtctagact acgatggtat ctaatcatct tcgatcccca gactttcgtt cttgatcaat 840 gaaaacatgc ttggtaaatg ccttcgcttt ggtgcgtctt tcgaaaatcc aagaatttca 900 cctctagctt cgaaatacgg atacccccaa ccgttcctat tcaccattac ctggacgagc 960 aaaccaatag ccacgcccca gtccctgtct tatcatcttg taacgaacgt accggcccta 1020 cggcctgctt ggagcactcc gatttcatta cagtgaacga ttcgatccga aaatggacca 1080 aaaaagtggc caggcacaag acgcacgcaa tggccgaagc caaagggcac ccgcacccca 1140 ccagacattc aactacgagc tttttaactg caacaacgtt tgtatagatt actggagctg 1200 gaattaccgc ggctgctggc accagacttg ccctccagtg gatcctcgat gagggtttta 1260 cattgctctc attccaatcg caaaacgcgg agcgcttcgc attggtattt ctcgtcacta 1320 cctcgcagag tctgcattgg gtaatttacg cgcctgctgc cttccttgga tgtggtagcc 1380 gtctctcagg ctccctctcc ggagtcgagc ccacactctc cggcacccgt tacggtcacc 1440 gtagtccaat acactaccgt cgacaactga tggggcagaa gctcacacga ttcaccgggc 1500 aaacacccag tccgcttagt tgctgtgaat caccaaaagc ttggcttgaa cccaacaaat 1560 gcagccccga aaggctctct tggcatgtat tgtttccaga attactgcaa gtatccatgt 1620 aaatggacca atcgttttgg tcacgactgt actaatgagc cgttcgcagc ttcaccgttt 1680 aatcgcttaa acttagacat gcatggctta atctttgaga cgaatctatg acagctggca 1740 ggatcaacca ggta 1754 <110> AVESTHAGEN LIMITED <120> DOCOSAHEXAENOIC ACID (DHA) PRODUCING THRAUSTOCHYTRID STRAIN-SC1 <160> 10 <170> KopatentIn 1.71 <210> 1 <211> 1547 <212> DNA <213> Thraustochtrid <220> <221> rRNA (222) (1) .. (1547) <400> 1 tacctggttg atcctgccag tagtcatatg ctcgtctcaa agattaagcc atgcatgtgt 60 aagtataagc gattgtactg tgagactgcg aacggctcat tatatcagta ataatttctt 120 cggtagtttc ttttatatgg atacctgcag taattctgga aataatacat gctgtaagag 180 ccctgtatgg ggctgcactt attagattga agccgatttt attggtgagt catgataatt 240 gagcagattg actattttgg tcgatgaatc gtttgagttt ctgccccatc agttgtcgac 300 ggtagtgtat tggactacgg tgactataac gggtgacgga gagttagggc tcgactccgg 360 agagggagcc tgagagacgg ctaccatatc caaggatagc agcaggcgcg taaattaccc 420 actgtggact ccacgaggta gtgacgagaa atatcgatgc gaagcgtgta tgcgttttgc 480 tatcggaatg agagcaatgt aaaaccctca tcgaggatca actggagggc aagtctggtg 540 ccagcagccg cggtaattcc agctccagaa gcatatgcta aagttgttgc agttaaaaag 600 ctcgtagttg aatttctggc atgggcgacc ggtgctttcc ctgaatgggg attgattgtc 660 tgtgttgcct tggccatctt tttcttttct ttttagggga gaaatctttc actgtaatca 720 aagcagagtg ttccaagcag gtcgtatgac cggtatgttt attatgggat gataagatag 780 gacttgggtg ctattttgtt ggtttgcacg cctgagtaat ggttaatagg aacagttggg 840 ggtattcgta tttaggagct agaggtgaaa ttcttggatt tccgaaagac gaactagagc 900 gaaggcattt accaagcatg ttttcattaa tcaagaacga aagtctgggg atcgaagatg 960 attagatacc atcgtagtct agaccgtaaa cgatgccgac ttgcgattgt tgggtgcttt 1020 tttaatgggc ctcagcagca gcacatgaga aatcaaagtc tttgggttcc ggggggagta 1080 tggtcgcaag gctgaaactt aaaggaattg acggaagggc accaccagga gtggagcctg 1140 cggcttaatt tgactcaaca cgggaaaact taccaggtcc agacataggt aggattgaca 1200 gattgagagc tctttcatga ttctatgggt ggtggtgcat ggccgttctt agttggtgga 1260 gtgatttgtc tggttaattc cgttaacgaa cgagacctcg gcctactaaa tagtgcgtgg 1320 tatggcaaac atagtacgtt tttaacttct tagagggaca tgtccggttt acgggcagga 1380 aagttcgagg caataacagg tctgtgatgc ccttagatgt tctgggccgc acgcgcgcta 1440 cactgatggg ttcatcgggt tttaatttca atttttggaa ttgagtgctt ggtcggaagg 1500 cctggctaat ccttggaacg ctcatcgtgc tggggctaga ttttgca 1547 <210> 2 <211> 1840 <212> DNA <213> Thraustochtrid <400> 2 gctccacgcg ttgggagctc tcccctatgg tcgacctgca ggcggccgcg aattcactag 60 tgatttacct ggttgatcct gccagtagcc ctacgctcgt ctcaaagact aagccatgca 120 tgtgtaagta taagcgaatt atactgtgaa actgcgaacg gctcattata tcagttataa 180 tcccttcggt agttccttta cacggatacc tgcagtaatt ctggaattaa tacgtgctgt 240 acgggcccga ctttcgggga gggccgcact tattaggtct aagccaacgt tattggtgag 300 tcatgataat tgagcagatc gcttttcgga gcgatgaatc gtttgagttt ctgccccatc 360 agttgtcgac ggtagggtat tggcctacgg tgactataac gggtgacggg gagttagggc 420 tcgactccgg agagggagcc tgagagacgg ctaccacatc caaggaaggc agcaggcgcg 480 taaattaccc aatgtggact ccacgaggta gtgacgagaa atatcaatgc ggggcgcttc 540 gcgtcttgct attggaatga gagcaatgta aaaccctcat cgaggatcaa ctggagggca 600 agtctggtgc cagcagccgc ggtaattcca gctccagaag cgtatgctaa agttgttgca 660 gttaaaaagc tcgtagttga atttctggtg tgggagccca ggcctcggtg cgaatgcgcc 720 ttgtactgct ttgcggctcc tttgccatcc tcgtttttcg taagaaaggc gtcattcact 780 gtaatcaaag cagagtgttc caagcaggcc gtagggccgg tatgtttatt atgggatgat 840 cagataggac tcgggtgcta ttttgttggt ttgcacatct gagtaatgat taataggaac 900 agtcgggggt atccgtattt aggagctaga ggtgaaattc ttggatttcc gaaagacgaa 960 ctacagcgaa ggcattacca agcatgtttt cattaatcaa gaacgaaagt ctggggatcg 1020 aagatgatta gataccatcg tagtctagac cgtaaacgat gccgacttgc gattgcgggt 1080 ggcttgtatt gggcctccgc agcagcacat gagaaatcaa agtctttggg ttccgggggg 1140 agtatggtcg caaggctgaa acttgaagga attgacggaa gggcaccacc aggagtggag 1200 cctgcggctt aatttgactc aacacgggaa aacttaccag gtccagacat agtaggattg 1260 acagattgag agctctttct tgattctatg ggtggtggtg catggccgtt cttagttggt 1320 ggagtgattt gtctggttaa ttccgttaac gaacgagacc tcggcctact aaatagccgg 1380 gcgtatggcg acatatgtgt ttgtggcttc ttagagggac atgttcggtt tacgagcagg 1440 aagttcgagg caataacagg tctgtgatgc ccttagatgt tctgggccgc acgcgcgcta 1500 cactgatggg ttcagcgggt cttgttgtga tttatcgcag cgttgctttg tcggaaggca 1560 tggctaatcc tttgaacgcc catcgtgctg gggctagatt tttgcaatta ttaatctcca 1620 acgaggaatt cctagtaaac gcaagtcatc agcttgcatt gaatacgtcc ctgccctttg 1680 tacacaccgc ccgtcgcacc taccgattga acggtccgat gaaaccatgg gactaccttt 1740 tgagcgtttg ttcgcgatgg aggtgggaac tcgggtgaat cttattgttt agaggaaggt 1800 gaagtcgtaa caaggtttcc gtaggtgaac ctgcggaagg 1840 <210> 3 <211> 1798 <212> DNA <213> Thraustochytrid <400> 3 tacctggttg atcctgccag taatcatacg cttgtctcaa agattaagcc atgcatgtct 60 aagtataaag gattatactc tgaaactgcg aacggctcat taaaccagta ctaacctaat 120 tgatgatgaa agttagatgg atacttgtgg caaatctaga accaatacat gcttgggggc 180 ccgactttat gggagggctg catttatttg aaccaaacca atagctcgtt tcggcgggtt 240 tcgtgtgggt gagtcagaat aagtaagcga accctttttt cggagagggt gaatcattcg 300 agtttctgcc ccatcagttg tcgacgacac ggtattgacc tgtcgtgact gtcacgggtg 360 acggagaatt agggttcgat tccggagagg gagcctgaga gacggctacc acatccaagg 420 aaggcagcag gcgcgtaaat tactcaatgt taaatggacg aagtagtgac gagaaataac 480 aatgtggagc gctttgcgtt ttacaattgt aatgagagca gattaaaggg gtcatcgagg 540 atccattgga gggcaagtct ggtgccagca gccgcggtaa ttccagctcc aatagcgtat 600 attaaagttg ttgcagttaa aaagctcgta gttgaagaag ggttgtcttt ggtttggttt 660 tcgtatgaga actgaacatg gcaacattcg tctgtttttg aaagcgggca gaccgtttac 720 tgtaaaaaaa ttagagtgtt caaagcgtga tgtatgttgt ttgaatatag tagtatggaa 780 taatgagata ggactttgtt gttattttgt tggtttgcat ggcaaggtaa tgattaatag 840 ggacagttgg gggtattcgt attacgatgt cagaggtgaa attcttggat tttcgaaaga 900 cgaacaactg cgaaagcatt taccaaggat gttttcatta atcaagaacg aaagtcaggg 960 gatcgaagat gattagatac catcgtagtc ttgaccgtaa acaatgccga cttgcgattg 1020 tgttcttctt gttttagaga gggatgcagc agcacatgag aaatcaaagt ctttgggttc 1080 cggggggagt atggtcgcaa ggctgaaact taaaggaatt gacggaaggg caccaccagg 1140 agtggagcct gcggcttaat ttgactcaac acggggaaac ttaccaggtc cggacatagg 1200 aaggattgac agattgagag ctctttcttg attctatggg tggtggtgca tggccgttct 1260 tagttggtgg agtgatttgt ctggttaatt ccgttaacga acgagaccac agcctactaa 1320 ataatggtgt ttttttgttt taggaaagac atgtcgtttc ttagagggac atttcgggtt 1380 taccggaagg gagtttgtgg caataacagg tctgtgatgc ccttagatgt tctgggccgc 1440 acgcgcgcta cactgatcgg ttcaacgagt gtttgcagtt tcgttttgag actgtgttcc 1500 ttggtaggaa tgcctgggta atcttttgaa cgccgatcgt gatggggcta gatttttgca 1560 attgttaatc tccaacgagg aattcctagt aaacgcaagt catcagcttg cattgattac 1620 gtccctgccc cttgtacaca ccgcccgtcg cacctaccga ttgaacgatc cggtgagacc 1680 ttgggattgc tcgattttct tttcacgagg agagtcagta gcaagaactt gagcaaacct 1740 tatcgtttag aggaaggtga agtcgtaaca aggtttccgt aggtgaacct gcggaagg 1798 <210> 4 <211> 1881 <212> DNA <213> Thraustochytrid <400> 4 acgcgttggg agctctccca tctggtcgac ctgcaggcgg ccgcgaattc actagtgatt 60 ccttccgcag gttcacctac ggaaaccttg ttacgacttc accttcctct aaacgataag 120 gtttactcaa cttttcccgt catcacacga acaaacgcaa aacaacagga tccgaagatt 180 tcaccggatc gttcaatcgg taggtgcgac gggcggtgtg tacaaagggc agggacgtaa 240 tcaatgcaag ctgatgactt gcgtttacta ggaattcctc gttggagatt agaattgcaa 300 aaatctagcc ccatcacgat ggacatttaa aagattgccc ggacattcct gtctaggacc 360 gtcattcatc gttaaataaa caacacatac tcgttgaatc catcagtgta gcgcgcgtgc 420 ggcccagaac atctaagggc atcacagacc tgttattgcc acaaacttcc ttccggtaaa 480 accgaaatgt ccctctaaga agtctcgcca ctatgtcacc atagcacgta ctatttagta 540 ggctgtggtc tcgttcgtta acggaattaa ccagacaaat cactccacca actaagaacg 600 gccatgcacc accacccata gaatcaagaa agagctctca atctgtcaat ccttcctatg 660 tctggacctg gtaagttttc ccgtgttgag tcaaattaag ccgcaggctc cactcctggt 720 ggtgcccttc cgtcaattcc tttaagtttc agccttgcga ccatactccc cccggaaccc 780 aaagactttg atttctcatg tgctgctgcc caagcctatt tcaaaaaaac atcggacaat 840 cgcaagtcgg catcgtttac ggttaagact acgatggtat ctaatcatct tcgatcccct 900 aactttcgtt cttgattaat gaaaacatcc ttggtaaatg cttttcgcag ttgttcgtct 960 tttgtaaatc caagaatttc acctctgaca accaaatacg aataccccca actgtccctg 1020 ttaatcatta ctttgatgtg caaaccaaca aaatagcaac aaagtcctat cttattattc 1080 catactactc tattccaacc atacgatttg cttgaaacac tctaattttt ttacagtaaa 1140 tgatgcccac ccacgcccaa aggcgcaaac gagcaaccat ggccagatca aacacaaaag 1200 ccactctcag cattcgccga aaagcctggt ccgtcctacc aatacttcaa ctacgagctt 1260 tttaactgca acaactttaa tacacgctac cggagctgga gttaccgcgg ctgctggcac 1320 cagacttgcc ctccagttga tcctcgatga gggttttaga ttgctctcat tacagaaaga 1380 gtaggcttag cccaccccac tgttatttct cgtcactact tcgtcgaatt gacattgagt 1440 aatttgcgcg cctgctgcct tccttggatg gggtagccgt ctctcaggct ccctctccgg 1500 aatcgaaccc tgattctccg tcacccgtaa caatcatggt aggccaatac cctaccatcg 1560 acaattgatg gggcagaaac ttgaatgaac atcgccggca caaacacctc tgcgattcgc 1620 ctagttacta tgaatcacca aaacaacccc gagaggtatt ggtttatatc taataaatgc 1680 agcccttccc aaaagtcggg ccctaaagca tgtattattt ctagatttgc cacaagtatc 1740 catctaatca aacattttca aagaaactat aactgtttta atgagccgtt cgcagtttca 1800 ccgtataaat cgcttatact tagacatgca tggcttagtc tttgagacaa gcgtgcgatt 1860 actggcagga tcaaccaggt a 1881 <210> 5 <211> 1795 <212> DNA <213> Thraustochytrids <400> 5 ccttccgcag gttcacctac ggaaaccttg ttacgacttc accttcctct aaacaataag 60 attcacccga gttctgcctc tgtccaaaaa tcaatccaaa cagaaacatc ccatggtttc 120 atcggaccgt tcaatcggta ggtgcgacgg gcggtgtgta caaagggcag ggacgtattc 180 aatgcaagct gatgacttgc gtttactagg aattcctcgt tggagattaa taattgcaaa 240 aatctagccc cagcacgatg agcgttccaa ggattagcca ggccttccga ccaagcactc 300 aattccaaaa attagaatta aaacccgatg aacccatcag tgtagcgcgc gtgcggccca 360 gaacatctaa gggcatcaca gacctgttat tgcctcgaac ttcctgcccg taaaccggac 420 atgtccctct aagaagtaaa aacgcactat gttgccatac cacgcactat ttagtaggcc 480 gaggtctcgt tcgttaacgg aattaaccag acaaatcact ccaccaacta agaacggcca 540 tgcaccacca cccatagaat catgaaagag ctctcaatct gtcaatccta cctatgtctg 600 gacctggtaa gttttcccgt gttgagtcaa attaagccgc aggctccact cctggtggtg 660 cccttccgtc aattccttta agtttcagcc ttgcgaccat actccccccg gaacccaaag 720 actttgattt ctcatgtgct gctgctgagg cccatataat aaagcaccca acaatcgcaa 780 gtcggcatcg tttacggtct agactacgat ggtatctaat catcttcgat ccccagactt 840 tcgttcttga ttaatgaaaa catgcttggt aaatgccttc gctctagttc gtctttcgga 900 aatccaagaa tttcacctct agctcctaaa tacgaatacc cccaactgtt cctattaacc 960 attactcagg cgtgcaaacc aacaaaatag cacccaagtc ctatcttatc atcccataat 1020 aaacataccg gtcatacgac ctgcttggaa cactctgctt tgattacagt gaaagatctc 1080 ataccaaaat agcatgagaa agatggccaa ggcaacacag acaatcaatc cccattcagg 1140 gaaagcaccg gtcgcccatg ccagaaattc aactacgagc tttttaactg caacaacttt 1200 agcatatgct tctggagctg gaattaccgc ggctgctggc accagacttg ccctccagtt 1260 gatcctcgat gagggtttta cattgctctc attccgatag caaaacgcat acacgcttcg 1320 catcgatatt tctcgtcact acctcgtggg gtccacagtg ggtaatttac gcgcctgctg 1380 ctatccttgg atatggtagc cgtctctcag gctccctctc cggagtcgag ccctaactct 1440 ccgtcacccg ttatagtcac cgtagtccaa tacactaccg tcgacaactg atggggcaga 1500 aactcaaacg attcatcgac aaaaaagtca atctgctcaa ttatcatgat tcaccaataa 1560 aatcggcttc aatctaataa gtgcagcccc atacagggct cttacagcat gtattatttc 1620 cagaattact gcaggtatcc atataaaaga aactaccgaa gaaattatta ctgatataat 1680 gagccgttcg cagtctcaca gtacaatcgc ttatacttac acatgcatgg cttaatcttt 1740 gagacgagca tatgactact ggcaggatca accaggtaaa tcgaattccc gcggc 1795 <210> 6 <211> 1779 <212> DNA <213> Thraustochytrid <220> <221> rRNA (222) (1) .. (1779) <400> 6 tacctggttg atcctgccag tagtcatatg ctcgtctcaa agattaagcc atgcatgtgt 60 aagtataagc gattgtactg tgagactgcg aacggctcat tatatcagta ataatttctt 120 cggtagtttc ttttatatgg atacctgcag taattctgga aataatacat gctgtaagag 180 ccctgtatgg ggctgcactt attagattga agccgatttt attggtgaat catgataatt 240 gagcagattg acttttttgt cgatgaatcg tttgagtttc tgccccatca gttgtcgacg 300 gtagtgtatt ggactacggt gactataacg ggtgacggag agttagggct cgactccgga 360 gagggagcct gagagacggc taccatatcc aaggatagca gcaggcgcgt aaattaccca 420 ctgtggaccc cacgaggtag tgacgagaaa tatcgatgcg aagcgtgtat gcgttttgct 480 atcggaatga gagcaatgta aaaccctcat cgaggatcaa ctggagggca agtctggtgc 540 cagcagccgc ggtaattcca gctccagaag catatgctaa agttgttgca gttaaaaagc 600 tcgtagttga atttctggca tgggcgaccg gtgctttccc tgaatgggga ttgattgtct 660 gtgttgcctt ggccatcttt ctcatgctat tttggtatga gatctttcac tgtaatcaaa 720 gcagagtgtt ccaagcaggt cgtatgaccg gtatgtttat tatgggatga taagatagga 780 cttgggtgct attttgttgg tttgcacgcc tgaataatgg ttaataggaa cagttggggg 840 tattcgtatt taggagctag aggtgaaatt cttggatttc cgaaagacga actagagcga 900 aggcatttac ccaagcatgt tttcattaat caagaacgaa agtctgggga tcgaagatga 960 ttagatacca tcgtagtcta gaccgtaaac gatgccgact tgcgattgtt gggtgcttta 1020 ttatatgggc ctcagcagca gcacatgaga aatcaaagtc tttgggttcc ggggggagta 1080 tggtcgcaag gctgaaactt aaaggaattg acggaagggc accaccagga gtggagcctg 1140 cggcttaatt tgactcaaca cgggaaaact taccaggtcc agacataggt aggattgaca 1200 gattgagagc tctttcatga ttctatgggt ggtggtgcat ggccgttctt agttggtgga 1260 gtgatttgtc tggttaattc cgttaacgaa cgagacctcg gcctactaaa tagtgcgtgg 1320 tatggcaaca tagtgcgttt ttacttctta gagggacatg tccggtttac gggcaggaag 1380 ttcgaggcaa taacaggtct gtgatgccct tagatgttct gggccgcacg cgcgctacac 1440 tgatgggttc atcgggtttt aattctaatt tttggaattg agtgcttggt cggaaggcct 1500 ggctaatcct tggaacgctc atcgtgctgg ggctagattt ttgcaattat taatctccaa 1560 cgaggaattc ctagtaaacg caagtcatca gcttgcattg aatacgtccc tgccctttgt 1620 acacaccgcc cgtcgcacct accgattgaa cggtccgatg aaaccatggg atgtttctgt 1680 ttggattgat ttttggacag aggcagaact cgggtgaatc ttattgttta gaggaaggtg 1740 aagtcgtaac aaggtttccg taggtgaacc tgcggaagg 1779 <210> 7 <211> 1803 <212> DNA <213> Thraustochytrid <220> <221> rRNA (222) (1) .. (1803) <400> 7 ccttccgcag gttcacctac ggaaaccttg ttacgacttc accttcctct aaacgttaag 60 gtttgctcaa ctttggccac gaactcctgt cgccaggaac aacgtgccct ccgaaggtct 120 caccggaacg ttcaatcggt aggtgcgacg ggcggtgtgt acaaagggca gggacgtaat 180 caatgcaagc tgatgacttg cgtttactag gaattcctcg ttggagatca ataattgcaa 240 agatctagcc ccatcacgac tggtattcac acgattaccc aaaccttccg gcccaggact 300 ctgtcataac aacaaaaaca acgtgctccc ccagtcagtg tagcgcgcgt gcggcccaga 360 acatctaagg gcatcacaga cctgttattg ccacaaactt ccctctggta aacccaaagt 420 gtccctctaa gaagtgccat accactgtgt cgccacaacg tgcactattt agtaggctgt 480 ggtctcgttc gttaacggaa tcaaccagac aaatcactcc accaactaag aacggccatg 540 caccaccacc catagaatca agaaagagct ctcaatctgt caatccttcc tatgtctgga 600 cctggtaagt ttccccgtgt tgagtcaaat taagccgcag gctccactcc tggtggtgcc 660 cttccgtcaa ttcctttaag tttcagcctt gcgaccatac tccccccgga acccaaagac 720 tttgatttct catgcgctgc cgcctcccgc ttcaagagcc gggagacaat cgcgagtcgg 780 catcgtttac ggtatggact acgatggtat ctaatcatct tcgatccccc atactttcgt 840 tcttgatcaa tgaaaacatc cttggtaaat gctttcgcag tcgttcggcc ttctagtaat 900 ccaagaattt cacctctgac actagaattc gaataccccc aactgtccct cttcatcatt 960 ccgccaacgc gcaaaccaac aaaataccgc caacggccta tctcatcatc cccatactat 1020 catattccaa tatcaacctt tgcttgaaac accctaattt ttttacagtg aaagaggcaa 1080 gtcttacaac ttgccagact ggcagcgcac gtgtcactgt tcggcaagca ttcacccgcc 1140 gacggacacg cacacaaacc atacgttcga ctacgagctt tttaactgca acaactttag 1200 tatacgctat tggagctgga attaccgcgg ctgctggcac cagacttgcc ctccaattga 1260 tccccgatgg gggttttaaa tcgctctcat ttcagtcgag atacgcacag cgcacctcac 1320 tgctatttct cgtcactact tcgttctgta aacttgcagt aatttacgcg cctgctgcct 1380 tccttggatg tggtagccgt ctctcaggct ccctctccgg actcgaaccc taattccccg 1440 tcacccgtaa cggtcacggg aggccaatac cctcccgtcg acaactgatg gggctgaaac 1500 ttgaatgatt catcgccgcc cgagggctat gcgattcgct caattactat gactcaccat 1560 cacaaggccc gagagcctac tggtttaggt ctaataagtg cggccctccc caaaagtcgg 1620 gcctccaagc acgtattgat tctagatttg ccacatgtat ccgtctacac aacgcacaat 1680 catgggaatt ataacagata taatgagccg ttcgcggttt cgctgtatta ccaactaata 1740 ctatcgcatg catggcttaa tctttaagac aagcgtatga ccgctggcag gatcaaccag 1800 gta 1803 <210> 8 <211> 1779 <212> DNA <213> Thraustochytrid <220> <221> rRNA (222) (1) .. (1779) <400> 8 tacctggttg atcctgccag tagtcatatg ctcgtctcaa agattaagcc atgcatgtgt 60 aagtataagc gattgtactg tgagactgcg aacggctcat tatatcagta ataatttctt 120 cggtagtttc ttttatatgg atacctgcag taattctgga aataatacat gctgtaagag 180 ccctgtatgg ggctgcactt attagattga agccgatttt attggtgaat catgataatt 240 gagcagattg acttttttgg tcgatgaatc gtttgagttt ctgccccatc agttgtcgac 300 ggtagtgtat tggactacgg tgactataac gggtgacgga gagttagggc tcgactccgg 360 agagggagcc tgagagacgg ctaccatatc caaggatagc agcaggcgcg taaattaccc 420 actgtggact ccacgaggta gtgacgagaa atatcgatgc gaagcgtgta tgcgttttgc 480 tatcggaatg agagcaatgt aaaaccctca tcgaggatca actggagggc aagtctggtg 540 ccagcagccg cggtaattcc agctccagaa gcatatgcta aagttgttgc agttaaaaag 600 ctcgtagttg aatttctggc atgggcgacc ggtgctttcc ctgaatgggg attgattgtc 660 tgtgttgcct tggccatctt tttcttttct ttttagggga gaaatctttc actgtaatca 720 aagcagagtg ttccaagcag gtcgtatgac cggtatgttt attatgggat gataagatag 780 gacttgggtg ctattttgtt ggtttgcacg cctgagtaat ggttaatagg aacagttggg 840 ggtattcgta tttaggagct agaggtgaaa ttcttggatt tccgaaagac gaactagagc 900 gaaggcattt accaagcatg ttttcattaa tcaagaacga aagtctgggg atcgaagatg 960 attagatacc atcgtagtct agaccgtaaa cgatgccgac ttgcgattgt tgggtgcttt 1020 tttatgggcc tcagcagcag cacatgagaa atcaaagtct ttgggttccg gggggagtat 1080 ggtcgcaagg ctgaaactta aaggaattga cggaagggca ccaccaggag tggagcctgc 1140 ggcttaattt gactcaacac gggaaaactt accaggtcca gacataggta ggattgacag 1200 attgagagct ctttcatgat actatgggtg gtggtgcatg gccgttctta gttggtggag 1260 tgatttgtct ggttaattcc gttaacgaac gagacctcgg cctactaaat agtgcgtggt 1320 atggcaacat agtacgtttt tacttcttag agggacatgt ccggtttacg ggcaggaagt 1380 tcgaggcaat aacaggtctg tgatgccctt agatgttctg ggccgcacgc gcgctacact 1440 gatgggttca tcgggtttta attctgtttt tatggaattg agtgcttggt cggaaggcct 1500 ggctaatcct tggaacgctc atcgtgctgg ggctagattt ttgcaattat taatctccaa 1560 cgaggaattc ctagtaaacg caagtcatca gcttgcattg aatacgtccc tgccctttgt 1620 acacaccgcc cgtcgcacct accgattgaa cggtccgatg aaaccatggg atgtttctgt 1680 ttggattgat ttttggacag aggcagaact cgggtgaatc ttattgttta gaggaaggtg 1740 aagtcgtaac aaggtttccg taggtgaacc tgcggaagg 1779 <210> 9 <211> 1793 <212> DNA <213> Thraustochytrid <400> 9 ccttccgcag gttcacctac gaaaaccttg ttacgacttc tccttcctct atgtgctgtg 60 gttcaccaaa ctttccactc ctcaaagaga gactcttatg aggaacggtc cgaattattc 120 accgaagcac acaatcggta ggagcgacgg gcggtgtgta caaagggcag ggacgtaatc 180 aatacgagat gttgactcgt gtttactagg aattcctcgt tgaagactaa taattgcaag 240 agtctatccc catcacgatg cattttaaag agattaccca acccttccag gcaaggaaat 300 acactcgttg aatgcatcag tgtagcgcgc gtgcagccca gaacatctaa gggcatcaca 360 gacctgttat tgcctcagac ttccttgcat tgaacataca aagtccctct aagaagctta 420 cccacatagc ctaggccatg tgtagctatt tagtaggctg aggtctcgtt cgttaacgga 480 attaaccaga caaatcactc caccaactaa gaacggccat gcaccaccac ccatagaatc 540 aggaaagagc tatcaatctg tcaatccttc caatgtctgg atctggtaag tttccccgtg 600 ttgagtcaaa ttaagccgca ggctccactc ctggtggtgc ccttccgtca attcctttta 660 agtttcagcc ttgcgaccat actcccccca gaacccaaag acttggattt ctcatcaagg 720 tgctgagaga gcaataatga cactcacccc aatctctagt tggcatggtt catggctaag 780 actaggacgg tatctaatcg tcttcgatcc cctaactttc cgttcttgat caaagaaaac 840 atccttggca aatgctttcg cattagtgtg tctttaacaa atccaagaat ttcacctctg 900 acagttaaat acaaatgccc ccaattgtcc ctatcaatcg ttgcgatggt tctcgaaacc 960 aataaaaaag atccaaagct caatttcatt attccatgct aatatattca aggcacatgc 1020 ctgcttgaaa cactctaatt tcctcaaagt aaaatagctg acaactatac gaccagcgaa 1080 ggccctatag tcatcaagac tcatgcacag aacatgtcag atactcaccc aggggatgga 1140 ccaacagcct ctatgcagaa attcaactac gagcttttta actgcaacaa ctttaatata 1200 cgctattgga gctggaatta ccgcggctgc tggcaccaga cttgccctcc aattgatact 1260 tgcaaagtgt tttaaattct gctcattcca attacaagac atggatgccc tgtattgtta 1320 tttcttgtca ctacctcctt gtgtcaagat tgggtaattt gcgcgcctgc tgccttcctt 1380 agatgtggta gccatttctc aggctccctc tccggaatcg aaccctaatt ctccgttacc 1440 cgtcattgcc atggtaggcc aataccctac catccaaagc tgataggtca gaaactcatg 1500 agatgcatca ccagaaagac catgtgattc atcctcttac catacttcat tatgatacca 1560 agcaagcttg gattggttct gaatttggta agcaccaccc tccaataagt tgggtgttga 1620 aacatgtatt ggctccagaa ttactgtagt tatccatata acgataatca tcaaatagat 1680 cattactgtt ataatgagcc attcgcagct taaccgtata aaggcataga ctcagacatg 1740 catggcttaa tctttgagac aagcatatga ctactggcag gatcaaccag gta 1793 <210> 10 <211> 1754 <212> DNA <213> thraustochytrid <400> 10 tccttccgca ggttcaccta cggaaacctt gttacgactt caccttcctc taaacgataa 60 gattcgtcga agttcaccgc gcgagccaag tcgccccaac tcgcgcagct cccgacgcct 120 catcggaccg ttcaatcggt aggtgcgacg ggcggtgtgt acaaagggca gggacgtatt 180 cgatgcgagc tgatgactcg cgtctactag gaattcctcg ttggagacca ataattgcaa 240 aggtctagcc ccagcacgac gagcgttcac aggattcgcc gccccctccg aggaagctca 300 gctccgaaaa gcaacacccg atgaacccgt cagtgtagcg cgcgtgcggc ccagaacatc 360 taagggcatc acagacctgt tattgcctcg aacttcccgc tcgtataccg aacatgtccc 420 tctaagaagc ctgcaccatg tcgccatgcc gcagcctagt tagtaggccg aggtctcgtt 480 cgttaacgga attaaccaga caaatcactc caccaactaa gaacggccat gcaccaccac 540 ccatagaatc atgaaagagc tctcaatctg tcaatcctac ctatgtctgg acctggtaag 600 tttccccgtg ttgagtcaaa ttaagccgca ggctccactc ctggtggtgc ccttccgtca 660 attcctttaa gtttcagcct tgcgaccata ctccccccgg aacccaaaga ctctggtttc 720 ccacacgctc ccgcagaagc tcacaagagc accctgcgat cgcgggtcgg catcgtttgc 780 ggtctagact acgatggtat ctaatcatct tcgatcccca gactttcgtt cttgatcaat 840 gaaaacatgc ttggtaaatg ccttcgcttt ggtgcgtctt tcgaaaatcc aagaatttca 900 cctctagctt cgaaatacgg atacccccaa ccgttcctat tcaccattac ctggacgagc 960 aaaccaatag ccacgcccca gtccctgtct tatcatcttg taacgaacgt accggcccta 1020 cggcctgctt ggagcactcc gatttcatta cagtgaacga ttcgatccga aaatggacca 1080 aaaaagtggc caggcacaag acgcacgcaa tggccgaagc caaagggcac ccgcacccca 1140 ccagacattc aactacgagc tttttaactg caacaacgtt tgtatagatt actggagctg 1200 gaattaccgc ggctgctggc accagacttg ccctccagtg gatcctcgat gagggtttta 1260 cattgctctc attccaatcg caaaacgcgg agcgcttcgc attggtattt ctcgtcacta 1320 cctcgcagag tctgcattgg gtaatttacg cgcctgctgc cttccttgga tgtggtagcc 1380 gtctctcagg ctccctctcc ggagtcgagc ccacactctc cggcacccgt tacggtcacc 1440 gtagtccaat acactaccgt cgacaactga tggggcagaa gctcacacga ttcaccgggc 1500 aaacacccag tccgcttagt tgctgtgaat caccaaaagc ttggcttgaa cccaacaaat 1560 gcagccccga aaggctctct tggcatgtat tgtttccaga attactgcaa gtatccatgt 1620 aaatggacca atcgttttgg tcacgactgt actaatgagc cgttcgcagc ttcaccgttt 1680 aatcgcttaa acttagacat gcatggctta atctttgaga cgaatctatg acagctggca 1740 ggatcaacca ggta 1754  

Claims (7)

도코사헥사엔산(DHA) 및 그들의 오메가-3-지방산/지방산 중간체를 생산하는 미생물에 있어서, 트라우스토키트리움(Thraustochytrium) 속을 포함하는 군으로부터 선택된 오메가-3-지방산 생산 미생물.An omega-3-fatty acid producing microorganism selected from the group comprising the Thraustochytrium genus for microorganisms producing docosahexaenoic acid (DHA) and their omega-3-fatty acid / fatty acid intermediates. 제1항에 있어서, 서열 번호 1로 표시되는 18S RNA 서열을 포함하는 오메가-3-지방산 생산 미생물.The omega-3-fatty acid producing microorganism according to claim 1, comprising an 18S RNA sequence represented by SEQ ID NO: 1. 제2항에 있어서, 도코사헥사엔산을 30% 이상 생산하는 오메가-3-지방산 생산 미생물.The omega-3-fatty acid producing microorganism according to claim 2, which produces at least 30% of docosahexaenoic acid. 제2항에 있어서, 도코사헥사엔산을 25% 이상 생산하는 메가-3-지방산 생산 미생물.The mega-3-fatty acid producing microorganism according to claim 2, which produces at least 25% of docosahexaenoic acid. 제2항에 있어서, 도코사헥사엔산을 20% 이상 생산하는 오메가-3-지방산 생산 미생물.The omega-3-fatty acid producing microorganism according to claim 2, which produces at least 20% of docosahexaenoic acid. 제2항에 있어서, 도코사헥사엔산을 15% 이상 생산하는 오메가-3-지방산 생산 미생물.The omega-3-fatty acid producing microorganism according to claim 2, which produces at least 15% of docosahexaenoic acid. 제2항에 있어서, 도코사헥사엔산을 10% 이상 생산하는 오메가-3-지방산 생산 미생물.3. The omega-3-fatty acid producing microorganism according to claim 2, which produces at least 10% of docosahexaenoic acid.

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