KR20110112541A - Cholesterol-lowered fermented meat products by use of lactobacilus and a method of producing the same - Google Patents
Cholesterol-lowered fermented meat products by use of lactobacilus and a method of producing the same Download PDFInfo
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Abstract
본 발명에 따라 김치로부터 콜레스테롤 저하효능을 가진 Lactobacillus plantarum NR74(KACC 91538P) 유산균 균주를 분리, 선별, 동정하여 발효용 스타터(starter culture)로 이용함으로써 저콜레스테롤 발효육 제품 및 그 제조방법이 제공된다.According to the present invention, a low cholesterol fermented meat product and a method of preparing the same are provided by separating, selecting, and identifying Lactobacillus plantarum NR74 (KACC 91538P) lactic acid bacteria strain having cholesterol lowering effect from kimchi as a starter culture for fermentation.
Description
본 발명은 김치로부터 콜레스테롤 저하효능을 가진 예컨대 Lactobacillus plantarum NR74와 같은 유산균 균주를 분리, 선별, 동정하여 발효용 스타터(starter culture)로 이용함으로써 저콜레스테롤 발효육 제품 및 그 제조방법을 제공하는 것이다. The present invention is to provide a low-cholesterol fermented meat product and method for producing the same by separating, selecting and identifying a lactic acid bacteria strain such as Lactobacillus plantarum NR74 having cholesterol lowering effect from kimchi as a starter culture for fermentation.
김치는 한국의 대표적 전통 발효식품으로서, 김치 발효에 관여하는 다양한 유산균 및 이것을 육류의 가공 등에 응용한 다양한 기술이 제안되어 온 바 있다.Kimchi is Korea's representative traditional fermented food, various lactic acid bacteria involved in the fermentation of kimchi and various techniques applied to the processing of meat has been proposed.
예를 들어, 한국특허출원 10-2000-0024426호에서는, 김치 발효에 관여하는 혐기성, 내염성의 30여 종의 유산균 중에서 20℃ 이상에서 잘 증식하여 젖산을 다량 생성하여 유해균의 생장, 증식을 억제하고 혐기 상태를 만들어주는 유산간균류 (Lactobacillus 속)의 우점적 증식환경(20℃이상)과, 저온(10℃이하)에서 우점적 증식하여 풍미에 관여하는 각종 유기산을 생성하는 유산구균류(Leuconostoc 속, Lactococcus 속)의 우점적 증식환경(10℃이하)의 발효환경(온도)차이를 이용하여 유산균 식이섬유 함유 발효육을 제조할 수 있는 제조공법이 개시되어 있다.For example, in Korean Patent Application No. 10-2000-0024426, among 30 kinds of anaerobic and flameproof lactic acid bacteria involved in kimchi fermentation, it proliferates well at 20 ° C or higher to generate a large amount of lactic acid to inhibit the growth and growth of harmful bacteria. Lactobacillus (Lactobacillus genus), which produces anaerobic conditions, predominantly grows above 20 ℃ and predominantly at low temperature (below 10 ℃) to produce various organic acids involved in flavor (Leuconostoc genus) , Lactococcus genus) is a manufacturing method for producing fermented meat containing lactobacillus dietary fiber using the difference in the fermentation environment (temperature) of the dominant growth environment (10 ℃ or less).
현대인들의 식습관에서는 육류 섭취가 많아지고 육류에 포함된 콜레스테롤의 체내 축적이 증가해온 추세인 바, 성인성 질환의 원인물질로 알려진 콜레스테롤의 증가가 국민건강을 위협하는 주요 원인으로 지적되어 오고 있다.In modern people's eating habits, meat consumption has been increasing and the accumulation of cholesterol in meat has been increasing. As a result, the increase in cholesterol, which is known as a cause of adult disease, has been pointed out as a major threat to national health.
전통 발효식품중 하나인 김치에는 다양한 유산균이 다량 함유되어 있고 김치의 발효작용에는 여러가지 유산균이 관여하며 그중에는 콜레스테롤을 분해 또는 저하시키는 종류의 유산균도 포함되어 있는 것으로 보고된 바 있다.Kimchi, one of the traditional fermented foods, contains a large amount of various lactic acid bacteria, and it has been reported that various kinds of lactic acid bacteria are involved in the fermentation of kimchi, and among them, lactic acid bacteria that decompose or lower cholesterol.
따라서, 본 발명의 목적은 유산균 발효에 의해 육류 본연의 풍미를 살리면서도 콜레스테롤 함유량을 현저히 저하시킨 건강지향적이며 기호성이 높은 발효육을 제공하는 것이다.Accordingly, it is an object of the present invention to provide a health-oriented and palatable fermented meat which significantly lowers cholesterol content while maintaining the natural flavor of meat by lactic acid bacteria fermentation.
본 발명의 다른 목적은 콜레스테롤을 분해하는 특성을 가지는 김치 유래의 유산균 및 이것을 포함하는 발효용 스타터를 제공하는 것이다.Another object of the present invention is to provide a kimchi-derived lactic acid bacteria having a property of degrading cholesterol and a starter for fermentation comprising the same.
본 발명의 또다른 목적은 콜레스테롤 함유량을 현저히 저하시킨 건강지향적이며 기호성이 높은 발효육을 제조하는 방법을 제공하는 것이다.It is still another object of the present invention to provide a method for producing health-oriented and high-fermentation fermented meat which significantly lowers cholesterol content.
상기와 같은 본 발명의 목적을 달성하기 위하여, 본 발명자들은 다양한 생김치 또는 숙성 김치로부터 채집한 김치 시료들로부터 김치 발효에 작용하는 수많은 유산균 등의 균주들을 분리, 선별하고 그 특성들에 따라 균주를 동정하여 콜레스테롤 저하작용을 나타내는 유산균 균주들만을 단리해내고, 이 균주들을 스타터 물질로 사용하여 소정의 조건에서 가공된 육류를 발효시킴으로써 콜레스테롤 함량을 현저하게 감소시킨 발효육 제품을 제조할 수 있게 되어 본 발명을 완성하게 되었다.In order to achieve the object of the present invention as described above, the inventors isolate and select a number of strains, such as lactic acid bacteria acting on kimchi fermentation from the kimchi samples collected from various raw kimchi or aged kimchi and identify the strains according to their characteristics To isolate only lactic acid bacteria strains exhibiting a cholesterol lowering effect, and ferment the processed meat under predetermined conditions using these strains as starter materials, thereby producing a fermented meat product which significantly reduced cholesterol content. To complete.
본 발명에 따라, 콜레스테롤을 분해하는 특성을 가지는 김치 유래의 유산균 및 이것을 포함하는 발효용 스타터를 사용하여 콜레스테롤 함유량을 현저히 저하시킨 건강지향적이며 기호성이 높은 발효육을 제공할 수 있게 되었다.According to the present invention, it is possible to provide a health-oriented, high-fermentation fermented meat that significantly lowers cholesterol content by using lactic acid bacteria derived from kimchi and cholesterol-derived fermentation starter.
도 1은 본 발명의 일실시예에 따른 콜레스테롤 저하 유산균주를 이용한 발효육 제품의 제조방법을 공정 단계별로 도시한 것이고,
도 2는 본 발명에 따라 김치에서 단리된 유산균의 in vitro 콜레스테롤 저하효과 시험 결과를 도시한 것이며,
도 3a 및 도 3b는 각각 본 발명에 따라 김치로부터 단리한 유산균을 사용하여 제조한 발효육을 섭식시킨 마우스 생체내 총 혈청 콜레스테롤 및 HDL 수준을 도시한 것이다.1 is a step-by-step diagram illustrating a method for preparing a fermented meat product using cholesterol-lowering lactic acid strains according to an embodiment of the present invention.
In Figure 2 is of the lactic acid bacteria isolated from kimchi in accordance with the invention In vitro cholesterol lowering effect test results are shown,
Figures 3a and 3b respectively show the total serum cholesterol and HDL levels in mice fed fermented meat prepared using lactic acid bacteria isolated from kimchi according to the present invention.
본 발명의 바람직한 일실시예에 따르면, 본 발명에 따라 제공되는 유산균의 발효작용에 의해 육류 본연의 풍미를 살리면서도 콜레스테롤 함유량을 현저히 저하시킨 건강지향적이며 기호성이 높은 발효육이 제공된다.According to a preferred embodiment of the present invention, by providing a fermentation action of the lactic acid bacteria provided in accordance with the present invention, while maintaining the natural flavor of meat while providing a health-oriented, high-fermentation fermented meat significantly reduced cholesterol content.
본 발명의 다른 바람직한 일실시예에 따르면, 콜레스테롤을 분해하는 특성을 가지는 김치 유래의 유산균 및 이것을 포함하는 발효용 스타터를 제공하는 것이다.According to another preferred embodiment of the present invention, it is to provide a lactic acid bacteria derived from kimchi and a fermentation starter comprising the same.
본 발명의 또다른 바람직한 일실시예에 따르면, 본 발명에 따라 제공되는 콜레스테롤 분해 특성을 가지는 김치 유래의 유산균 및 이것을 포함하는 발효용 스타터를 사용하여, 콜레스테롤 함유량을 현저히 저하시킨 건강지향적이며 기호성이 높은 발효육을 제조하는 방법이 제공된다.According to another preferred embodiment of the present invention, by using a lactic acid bacteria derived from kimchi having cholesterol decomposition properties and a fermentation starter comprising the same, a health-oriented and high palatability significantly reduced cholesterol content A method for producing fermented meat is provided.
본 발명에 따른 발효육의 제조방법은, 김치로부터 유산균주 Lactobacillus plantarum NR74를 선별, 분리하여 증폭시키는 단계; 원료육을 입자크기가 3∼5mm로 세절 및 혼합하고 Lactobacillus plantarum NR74를 포함하는 유산균 스타터 및 첨가 재료를 첨가하여 염지육을 제공하는 단계; 염지육을 케이징에 충전하고 습도 80 내지 95%, 온도 15~22℃에서 2일 내지 21일 훈연 및 발효시키는 단계; 및 발효된 유산균 발효육을 제공하는 단계를 포함한다.Fermented meat production method according to the present invention, the step of selecting, separating and amplifying the lactic acid strain Lactobacillus plantarum NR74 from kimchi; Cut and mix the raw meat with the particle size of 3 ~ 5mm and Lactobacillus plantarum Adding the lactic acid bacteria starter comprising NR74 and the additive material to provide salted meat; Filling the salted meat in the casing and smoking and fermenting for 2 to 21 days at a humidity of 80 to 95%, a temperature of 15 to 22 ℃; And providing the fermented lactic acid bacteria fermented meat.
본 발명자들의 상기 균주를 동정하기 위해 16S rRNA 염기서열을 자동 염기서열 분석기로 분석한 결과, 상기 균주는 락토바실러스 사케이 및 락토바실러스 플란타룸으로 동정되었다. 본 발명자들은 상기 균주를 '락토바실러스 플란타룸 NR74'로 명명하고, 이를 2010년 3월 12일자로 국립농업과학원 농업유전자원센터(KACC)에 각각 기탁하였다(수탁번호 : KACC 91538P).In order to identify the strains of the present inventors, 16S rRNA sequences were analyzed by an automatic sequence analyzer, and the strains were identified as Lactobacillus sakei and Lactobacillus plantarum. The inventors named the strain 'Lactobacillus plantarum NR74', and deposited it on March 12, 2010 at the National Institute of Agricultural Science (KACC) (Accession Number: KACC 91538P).
이하, 본 발명을 실시예에 의해 상세하게 설명할 것인 바, 하기 실시예는 본 발명을 더욱 구체적으로 예시하기 위해 제공되는 것일 뿐, 본 발명의 사상과 범위가 하기 실시예에 의해 한정되는 것은 아니다.
Hereinafter, the present invention will be described in detail by way of examples, the following examples are provided only to illustrate the present invention in more detail, and the spirit and scope of the present invention are limited by the following examples. no.
실시예Example
재료 및 방법Materials and methods
1. 김치로부터 유산균 분리1. Isolation of Lactic Acid Bacteria from Kimchi
(1)전통 발효 식품인 김치에서 유산균을 분리 및 선별하였다(1) Lactobacillus was isolated and screened from kimchi, a traditional fermented food
10g의 김치시료를 90ml 멸균수에 넣고 분쇄기(stomacher)에서 균질화 (homogenizing)를 수행하고, 각 시료에서 멸균수 106-107/ml취하여 10배 희석하였다. 희석시킨 시료를 Agar플레이트(deman, Rogosa, Sharpe(MRS:Difco: U.S.A))에 넣고 20℃로 48시간동안 인큐베이션하였다.10 g of kimchi sample was added to 90 ml of sterile water, homogenized (homogenizing) in a grinder, and each sample was diluted 10-fold by taking 10 6 -10 7 / ml of sterile water. Diluted samples were placed in Agar plates (deman, Rogosa, Sharpe (MRS: Difco: USA)) and incubated at 20 ° C. for 48 hours.
각각 모양과 군락상태가 다른 균주들을 선택해 분리하고, 분리된 균주들을 배양액에 접종하여 배양한 후 -70℃에 보관하여 발효용 스타터 물질로 사용하였다. Strains of different shapes and colonies were selected and separated, and the isolated strains were inoculated in a culture solution and incubated at -70 ° C to be used as a starter material for fermentation.
분리한 미생물 중 유산균을 선별하기 위해 그람 염색법으로 염색된 미생물을 현미경 관찰하였다. 0.3% (v/v) H2O2(Junsei,Japan)용액을 사용하는 촉매 테스트 (catalase test)를 통하여 식품에서 분리한 미생물중 유산균을 선별하였다.Microorganisms stained by Gram staining were examined under a microscope to select lactic acid bacteria among the isolated microorganisms. Lactic acid bacteria were isolated from microorganisms isolated from foods through a catalase test using 0.3% (v / v) H 2 O 2 (Junsei, Japan) solution.
(2) 선별된 유산균의 동정(2) Identification of selected lactic acid bacteria
선별된 유산균의 동정을 위해 16s rDNA 서열분석기(sequencing)를 이용하였다. 16s rDNA sequencing was used to identify the selected lactic acid bacteria.
선별 균주의 16S rDNA는 Escherichia coli의 16S rDNA 유전자의 8-27과 1511-1491 위치에 각각 상응하는 2개의 프라이머, 즉 16S seq fw (5'-AGA GTT TGA TCM TGG CTC AG-3') 과 16S seq rev (5'-GGN TAC CTT GTT ACG ACT TC-3')를 사용하여 PCR로 증폭하였다. PCR 반응은 rep-PCR에서 기술된 바와 같이 반응혼합물을 조성하여 35 사이클 (94℃에서 1분, 56℃에서 1분, 72℃에서 2분)을 실행하였다. The 16S rDNA of the selected strain was composed of two primers corresponding to positions 8-27 and 1511-1491 of the 16S rDNA gene of Escherichia coli, namely 16S seq fw (5'-AGA GTT TGA TCM TGG CTC AG-3 ') and 16S. Amplification by PCR using seq rev (5'-GGN TAC CTT GTT ACG ACT TC-3 '). The PCR reaction was carried out 35 cycles (1 minute at 94 ° C., 1 minute at 56 ° C., 2 minutes at 72 ° C.) with the reaction mixture as described for rep-PCR.
PCR 반응물은 Quantum prep-PCR Clean Column(Biorad, Germany)로 정제하며 GATC Biotech (GATC, Konstanz, Germany)에서 양방향으로 서열분석(sequencing)을 수행한 후, 분석된 서열을 Blast에 입력하여 선별된 유산균을 동정하였다.The PCR reactions were purified by Quantum prep-PCR Clean Column (Biorad, Germany), sequencing in both directions at GATC Biotech (GATC, Konstanz, Germany), and the selected lactic acid bacteria were selected by inputting the analyzed sequences into Blast. Was identified.
2. 콜레스테롤(Cholesterol) 저하 유산균의 특성 조사2. Characterization of cholesterol lowering lactic acid bacteria
(1) 김치에서 콜레스테롤 저하 기능성 미생물 선별(1) Cholesterol-lowering functional microorganism screening in kimchi
10g의 김치시료를 90ml 멸균수에 넣고 분쇄기(stomacher)에서 균질화(homogenizing)를 수행하고, 각 시료에서 멸균수 106-107/ml취하여 10배 희석하였다. 희석시킨 시료를 Agar플레이트(deman, Rogosa, Sharpe(MRS:Difco: U.S.A))에 넣고 20℃로 48시간동안 인큐베이션하였다.10 g of kimchi sample was added to 90 ml of sterile water, homogenized (homogenizing) in a grinder, and each sample was diluted 10-fold by taking 10 6 -10 7 / ml of sterile water. Diluted samples were placed in Agar plates (deman, Rogosa, Sharpe (MRS: Difco: USA)) and incubated at 20 ° C. for 48 hours.
각각 모양과 군락상태가 다른 균주들을 선택해 분리하고, 분리된 균주들을 배양액에 접종하여 배양한 후 -70℃에 보관하여 발효용 스타터 물질로 사용하였다. Strains of different shapes and colonies were selected and separated, and the isolated strains were inoculated in a culture solution and incubated at -70 ° C to be used as a starter material for fermentation.
분리한 미생물 중 유산균을 선별하기 위해 그람 염색법으로 염색된 미생물을 현미경 관찰하였다. 0.3% (v/v) H2O2(Junsei,Japan)용액을 사용하는 촉매 테스트 (catalase test)를 통하여 식품에서 분리한 미생물중 유산균을 선별하였다.Microorganisms stained by Gram staining were examined under a microscope to select lactic acid bacteria among the isolated microorganisms. Lactic acid bacteria were isolated from microorganisms isolated from foods through a catalase test using 0.3% (v / v) H 2 O 2 (Junsei, Japan) solution.
(2)내담즙성 및 내산성 조사(2) Bile resistance and acid resistance investigation
유산균의 내산성은 배양액의 초기 pH를 2∼8까지 조절하여 밤새배양(overnight culture)한 유산균을 1% 비율로(v/v) 접종하여 균주의 생육을 측정하였다. 유산균의 내담즙성은 0.3%∼1%농도의 oxgall을 배양액에 첨가한 후, 밤새배양(overnight culture)한 유산균을 1% 비율로(v/v) 접종하여 37℃에서 배양함으로써 균주의 생육을 측정하였다.Acid resistance of the lactic acid bacteria was measured by inoculating the initial pH of the culture medium to 2 to 8 and inoculated at 1% rate (v / v) of the lactic acid bacteria cultured overnight (overnight culture) was measured. The bile resistance of lactic acid bacteria was measured by adding oxgall at a concentration of 0.3% to 1% to the culture medium, and then inoculating 1% of the lactic acid bacteria cultured overnight (v / v) and incubating at 37 ° C. It was.
(3)담즙산 분해효소 생성 실험(3) Bile acid degrading enzyme production experiment
담즙산 분해효소를 생성하는 유산균이 혈중 콜레스테롤을 감소시킨다는 국내외 다수 연구들을 바탕으로 유산균을 18시간 동안 액체 배지에 배양하였다. Lactic acid bacteria were cultured in a liquid medium for 18 hours based on a number of studies at home and abroad, which showed that lactic acid bacteria producing bile acid enzymes reduce blood cholesterol.
유산균을 0.5% (v/v) 타우로데옥시콜린산 [sodium salt of taurodeoxycholic acid (TDCA)](Sigma, St. Louis, U.S.A) 나트륨염과 0.37g/l CaCl2 (Sigma,St.Louis,U.S.A)이 포함된 MRS 고체 배지 위에 놓은 6mm 필터 디스크(filter disk) (Whatman, England)에 접종하였다.Lactic acid bacteria 0.5% (v / v) sodium salt of taurodeoxycholic acid (TDCA) (Sigma, St. Louis, USA) sodium salt and 0.37 g / l CaCl 2 6 mm filter disk (Whatman, England) was placed on MRS solid medium containing (Sigma, St. Louis, USA).
접종된 배지를 혐기적으로 37℃에서 72시간 동안 배양한 후 디스크 주위에 형성된 침전 구획을 측정하였다.The inoculated medium was anaerobicly incubated at 37 ° C. for 72 hours and then the precipitation sections formed around the discs were measured.
3. 균주의 콜레스테롤 저하 효능의 측정과 선별3. Measurement and Screening of Cholesterol Lowering Efficacy of Strains
(1) 미생물의 콜레스테롤 저하 측정과 선별(1) Measurement and Screening of Cholesterol Lowering in Microorganisms
in vitro 실험을 통하여 콜레스테롤 저하효과를 갖는 유산균을 선별하기 위해, MRS 배지에 oxgall의 농도를 0%와 0.3%로 조절하여 첨가하고, 가용성 콜레스테롤을 1 mg/mL의 농도로 첨가하였다.In order to select lactic acid bacteria having cholesterol-lowering effects through in vitro experiments, oxgall concentration was adjusted to 0% and 0.3% in MRS medium, and soluble cholesterol was added at a concentration of 1 mg / mL.
이 배지에 밤새 배양(overnight culture)한 미생물을 1% 농도로 접종하여 접종하지 않은 대조군과 함께 37℃에서 24시간 배양하였다. 배지중의 남아있는 콜레스테롤의 함량을 정량하기 위하여 미생물 배양액을 5000g에서 10분간 원심 분리하여 그 상등액을 취하여 앞서 기술한 Rudel and Morris (1973)의 방법으로 측정하였다.Overnight culture microorganisms were inoculated at 1% concentration and cultured at 37 ° C. for 24 hours with the uninoculated controls. In order to quantify the amount of cholesterol remaining in the medium, the microbial culture was centrifuged at 5000 g for 10 minutes, and the supernatant was taken and measured by the method of Rudel and Morris (1973) described above.
(2) 고콜레스테롤 유도 마우스에 대한 선별 유산균 섭취 효과 조사(2) Investigation of the Effect of Selected Lactic Acid Bacteria on High Cholesterol-Induced Mice
선행연구에서 콜레스테롤 저하 효과가 가장 뛰어난 Lactobacillus plantarum NR74을 선별하여 마우스에 고지방식(HFD)과 함께 먹여 김치유산균의 마우스 혈중 콜레스테롤 저하효과를 확인하였다. Lactobacillus has the highest cholesterol-lowering effect in previous studies The screening plantarum NR74 mice fed with high fat (HFD) was confirmed in mouse serum cholesterol lowering effects of kimchi lactic acid bacteria.
마우스는 섭식에 의한 고콜레스테롤을 유도하기 위하여 HFD에 콜레스테롤 1%을 섞어 투여하였으며, ND 그룹을 대조군(control), HFD 그룹을 음성대조군(negative control)으로 활용하여, Lactobacillus plantarum NR74을 HFD에 섞어 먹였을 때 마우스의 혈중 콜레스테롤 저하효과를 확인하였다. Mice were administered with a mixture of 1% cholesterol in HFD to induce high cholesterol by feeding, and Lactobacillus by using ND group as a control and HFD group as a negative control. When the plantarum NR74 was mixed with HFD and fed to the blood cholesterol lowering effect of the mouse was confirmed.
마우스는 그룹당 8마리 나누었고, 4주령 C57BL/6 마우스 수컷을 사용하여 1주 안정화 기간을 주고, 4주 동안 식이에 의한 고콜레스테롤을 유도하여 김치유산균의 혈중 콜레스테롤 저하효과를 확인하였다. 혈중 콜레스테롤 정량은 콜레스테롤 키트를 사용하여 수행하며, 콜레스테롤 총량과 고밀도 지방단백질(High Density Lipoprotein)을 정량하였다.The mice were divided into 8 groups, and 4 week old males were given a stabilization period using C57BL / 6 male males, and high cholesterol was induced by diet for 4 weeks. Cholesterol quantification was performed using a cholesterol kit, and the total amount of cholesterol and high density lipoprotein were quantified.
4. 발효육 제품의 제조4. Preparation of Fermented Meat Products
(1)재료 및 방법(1) Materials and Methods
돼지고기는 국내산 돈육으로 도축후 48시간이 경과된 약 pH5.8 정도의 뒷다리 적육부위와 등 지방을 정육면체(50㎤)로 자른 후 선모충(Trichinella spiralis)을 불활성화시키기 위하여 -20℃ 냉동고에서 1주간 동결시킨 후 사용하였다.Cut the pork hind enemy thick above the extent of about pH5.8 to 48 hours after slaughtering domestic pork fat, such as elapsed cube (50㎤) trichina (Trichinella spiralis ) was used after freezing for 1 week in a -20 ℃ freezer to inactivate.
미생물은 김치에서 분리된 Lactobacillus plantarum NR74를 사용하였다.The microorganism is Lactobacillus isolated from Kimchi plantarum NR74 was used.
소금은 일반적인 순도 99.9%의 정제염을 사용하며, 발색제는 21일간의 숙성과정을 고려하여 질산염과 아질산염의 1:2의 비율로 혼합하여 사용하였다. 숙성기간 동안의 변색 방지와 지방산화를 억제하기 위하여 Vit-C를 사용하며, 향신료는 기본적인 후추와 코리안더 천연분말을 사용하였다.Salt was used as a general purity of 99.9% refined salt, and the coloring agent was used by mixing 1: 2 ratio of nitrate and nitrite in consideration of the aging process for 21 days. Vit-C is used to prevent discoloration and fatty acidization during the ripening period, and the basic pepper and coriander natural powder are used for spices.
케이싱은 직경 38mm의 통기성 섬유 케이징(Fibrous casing)을 30℃∼40℃의 온수에서 약 30분간 수화시킨 후 표면 물기를 제거하여 사용하였다.The casing was used by hydrating a breathable fibrous casing having a diameter of 38 mm in hot water at 30 ° C. to 40 ° C. for about 30 minutes, and then removing surface moisture.
(2)발효육 제품의 제조(2) production of fermented meat products
발효육 제품의 재료 배합비는 표 1에 나타낸 바와 같다. 동결된 원료육을 절단 작업이 용이한 상태가 되도록 해동시킨후(표면 2/3 결빙), 준비된 첨가물과 함께, 콜레스테롤 저하 효능 유산균 Lactobacillus plantarum NR74을 절단장치내에 투입하여 (Cutting 1단으로) 원료육의 입자크기가 3∼5mm정도 될 때까지 세절 및 혼합하여 염지육을 완성하며, 절단 수행후 절단육 온도는 4℃정도를 유지하였다.The ingredient ratio of the fermented meat product is shown in Table 1. After thawing the frozen raw meat to facilitate the cutting operation (2/3 freezing surface), together with the prepared additives, cholesterol lowering effect Lactobacillus plantarum NR74 was put into the cutting device (cutting stage 1) until the grain size of the raw meat was 3 ~ 5mm, and the salted meat was completed by mixing. After cutting, the cut meat temperature was maintained at about 4 ℃.
충전방법은 진공 충전기를 이용하여 발효육 제품내에 기포가 형성되지 않도록 하여 섬유 케이징 (Fibrous casing)에 충전한 후 참나무 훈연칩을 사용하여 냉훈법(30℃이하)으로 5시간 훈연을 실시하였다.Filling method was used in a vacuum charger to prevent bubbles from forming in the fermented meat product was filled in a fiber casing (Fibrous casing) and then smoked for 5 hours by cold smoking method (below 30 ℃) using oak smoked chips.
숙성 및 건조를 표 2와 같은 조건에서 실시하여 21일간 숙성과정을 거친 발효육 제품을 제조하였다.
Aging and drying were carried out under the conditions shown in Table 2 to prepare a fermented meat product after 21 days of aging.
5. 발효육 제품의 콜레스테롤 함량 검사5. Cholesterol Content Test of Fermented Meat Products
발효육 제품의 콜레스테롤 함량 검사를 실시하였다.Cholesterol content test of fermented meat product was performed.
도 2는 본 발명에 따라 김치에서 단리된 유산균의 in vitro 콜레스테롤 저하효과 시험 결과를 도시한 것이다. 도시된 바와 같이, thio-브로쓰를 사용한 대조군에 비해, 본 발명의 콜레스테롤 저하 효능 유산균으로 처리한 그룹에서 Lactobacillus plantarum NR74는 약 51.3%의 감소효과를 나타내었다.
In Figure 2 is of the lactic acid bacteria isolated from kimchi in accordance with the invention Results of in vitro cholesterol lowering effect test are shown. As shown, Lactobacillus plantarum NR74 showed a reduction of about 51.3% in the group treated with the cholesterol lowering effect lactic acid bacterium of the present invention, compared to the control using thio-broth.
6. 발효 육제품을 섭취한 쥐에서 콜레스테롤 저하 효과 검사6. Screening of Cholesterol-lowering Effects in Rats Fed with Fermented Meat Products
도 3a 및 도 3b는 각각 본 발명에 따라 김치로부터 단리한 유산균을 사용하여 제조한 발효육을 섭식시킨 마우스 생체내 총 혈청 콜레스테롤 및 HDL 수준을 도시한 것이다. Figures 3a and 3b respectively show the total serum cholesterol and HDL levels in mice fed fermented meat prepared using lactic acid bacteria isolated from kimchi according to the present invention.
Lactobacillus plantarum NR74를 제공한 마우스 그룹은 HFD를 제공한 그룹에 비하여 약 20%정도 총 콜레스테롤 수준이 감소하였으며, 그에 비해 고밀도 지방단백질(high density lipoprotein)의 양은 HFD 그룹에 비해 약 25%정도 증가하는 것을 확인할 수 있었다.
Lactobacillus The group of mice that received plantarum NR74 reduced total cholesterol levels by about 20% compared to the group that received HFD, whereas the amount of high density lipoproteins increased by about 25% compared to the HFD group. Could.
연구결과Results
1. 김치 유산균의 분리 및 동정1. Isolation and Identification of Kimchi Lactobacillus
여러 김치로부터 33개 균이 분리되었다. 27가지의 균이 카탈라아제 음성, 그람 양성으로 유산균이었고, 그 중 11개는 가스(gas)를 생성하지 않는 막대형상(rod shape) 간균들이었다. 김치에서 선별된 11가지 균중 1개 균주는 Lactobacillus plantarum으로 동정되었다.33 bacteria were isolated from various kimchi. Twenty seven bacteria were catalase-negative, gram-positive, and eleven of them were rod-shaped rods that did not produce gas. One of 11 strains selected from Kimchi was identified as Lactobacillus plantarum .
2. 콜레스테롤 저하 유산균의 특성 조사2. Characterization of cholesterol-lowering lactic acid bacteria
BSH 활성은 형성되는 존(zone)을 측정하여 확인할 수 있는데, 1종류의 Lactobacillus plantarum 균주가 선행연구 3에서 BSH 침전 구획(precipitation zone)을 형성하였다. 이는 담즙산 분해효소에 의한 콜레스테롤을 저하시켜 줄 수 있는 것을 나타내며, 김치로부터 분리한 12종류의 균주중 1종류의 Lactobacillus plantarum 균주는 콜레스테롤을 저하키는 기능성을 갖추었다.BSH activity can be confirmed by measuring the zones formed. One type of Lactobacillus plantarum strain formed the BSH precipitation zone in the previous study 3. This indicates that it can lower cholesterol caused by bile acid degrading enzyme, and one type of Lactobacillus among 12 strains isolated from kimchi plantarum strains have the ability to lower cholesterol.
3. 균주의 콜레스테롤 저하 효능의 측정과 선별3. Measurement and Screening of Cholesterol Lowering Efficacy of Strains
(1) 미생물의 콜레스테롤 저하 측정과 선별 (1) Measurement and Screening of Cholesterol Lowering in Microorganisms
- 콜레스테롤 동화(assimilation)시험에 의해 정량한 콜레스테롤 양이 따이오-브로쓰(Thio-broth) 대조군보다 감소하였다는 것은 유산균에 의해 콜레스테롤이 분해되었다는 것을 의미하는데, 40% 이상의 콜레스테롤 감소 효과를 나타내었고, Lactobacillus plantarum NR74는 약 51.3%의 콜레스테롤 감소효과를 보였다. 김치에서 분리한 본 발명의 유산균 Lactobacillus plantarum NR74는 콜레스테롤 저하효과를 가짐을 확인하였다.-The decrease in the amount of cholesterol quantified by cholesterol assimilation test, compared to the Thio-broth control group, means that cholesterol was degraded by lactic acid bacteria. , Lactobacillus plantarum NR74 decreased cholesterol by 51.3%. Lactobacillus Lactobacillus of the Invention Isolated from Kimchi plantarum NR74 has been shown to have a cholesterol-lowering effect.
(2) 선별된 유산균의 섭취가 고 콜레스테롤 유도 마우스(mouse)의 혈중 콜레스테롤에 미치는 영향 조사 (2) Study on the Effect of Selected Lactic Acid Bacteria on Blood Cholesterol in High Cholesterol-induced Mouse
Lactobacillus plantarum NR74를 제공한 마우스 그룹은 HFD를 제공한 그룹에 비하여 약 20% 정도 총 콜레스테롤 수준이 감소하였으며, 그에 비해 고밀도 지방단백질(high density lipoprotein)의 양은 HFD 그룹에 비해 약 25%정도 증가하는 것을 확인할 수 있었다. 이는 Lactobacillus plantarum NR74이 혈중 콜레스테롤 농도를 낮추는 효과를 가지고 있고, 저하된 콜레스테롤 성분중 HDL의 함량은 높이고, LDL과 TG의 함량을 낮추는 효과를 예상해 볼 수 있다. 이는 김치 유산균이 동맥경화를 예방할 수 있는 지질대사와 관련이 있음을 동물실험(in vivo study)를 통해 예상해 볼 수 있으며, HDL의 함량을 높임으로써 포말세포(foam cell) 형성을 억제시킬 수 있을 것으로 예상된다. Lactobacillus The group of mice that received plantarum NR74 reduced total cholesterol levels by about 20% compared to the group that received HFD, whereas the amount of high density lipoproteins increased by about 25% compared to the HFD group. Could. This is Lactobacillus plantarum NR74 has the effect of lowering the cholesterol level in the blood, and the effect of lowering the cholesterol content of HDL and lowering the content of LDL and TG can be expected. This can be predicted through in vivo studies that kimchi lactic acid bacteria are associated with lipid metabolism to prevent atherosclerosis, and it is possible to suppress foam cell formation by increasing HDL content. It is expected.
본 발명에 따른 상기 유산균 균주 및 스타터 물질이 제공됨에 따라 본 발명의 유산균 발효에 따른 발효육 제조방법으로 육류의 대량 처리공정이 가능하게 되었으며, 이에 따라 육류 본연의 풍미를 살리면서도 콜레스테롤 함유량을 현저히 저하시킨 건강지향적이며 기호성이 높은 저콜레스테롤 발효육을 제공하는 산업적 적용이 가능하게 되었다. As the lactic acid bacteria strain and the starter material according to the present invention are provided, a process for mass-processing meat is possible by the fermented meat manufacturing method according to the lactic acid bacteria fermentation of the present invention, thereby significantly lowering the cholesterol content while maintaining the natural flavor of meat. Industrial applications have been made to provide low-cholesterol fermented meat that is health-oriented and palatable.
Claims (4)
김치로부터 유산균주 Lactobacillus plantarum NR74를 선별, 분리하여 증폭시키는 단계;
원료육을 입자크기가 3∼5mm로 세절 및 혼합하고 Lactobacillus plantarum NR74를 포함하는 유산균 스타터 및 첨가 재료를 첨가하여 염지육을 제공하는 단계;
염지육을 케이징에 충전하고 습도 80 내지 95%, 온도 15~22℃에서 2일 내지 21일 훈연 및 발효시키는 단계; 및
발효된 유산균 발효육을 제공하는 단계
를 포함하는 것을 특징으로 하는 저콜레스테롤 발효육의 제조방법.As a method of producing fermented meat according to claim 4,
Lactobacillus Lactobacillus from Kimchi plantarum Screening, isolating and amplifying NR74;
Cut and mix the raw meat with the particle size of 3 ~ 5mm and Lactobacillus plantarum Adding the lactic acid bacteria starter comprising NR74 and the additive material to provide salted meat;
Filling the salted meat in the casing and smoking and fermenting for 2 to 21 days at a humidity of 80 to 95%, a temperature of 15 to 22 ℃; And
Providing fermented lactic acid bacteria fermented meat
Low cholesterol fermented meat manufacturing method comprising a.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101402914B1 (en) * | 2014-01-08 | 2014-06-02 | 최재일 | Fermented meat |
| CN110760456A (en) * | 2019-07-09 | 2020-02-07 | 怀化学院 | Lactobacillus plantarum La1 for degrading cholesterol and application thereof |
| KR20210158652A (en) | 2020-06-24 | 2021-12-31 | 주식회사 리얼푸드플랫폼 | Method for manufacturing of meat processing product using Vegetable Lactobacillus |
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2010
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101402914B1 (en) * | 2014-01-08 | 2014-06-02 | 최재일 | Fermented meat |
| CN110760456A (en) * | 2019-07-09 | 2020-02-07 | 怀化学院 | Lactobacillus plantarum La1 for degrading cholesterol and application thereof |
| CN110760456B (en) * | 2019-07-09 | 2022-03-22 | 怀化学院 | Lactobacillus plantarum La1 for degrading cholesterol and application thereof |
| KR20210158652A (en) | 2020-06-24 | 2021-12-31 | 주식회사 리얼푸드플랫폼 | Method for manufacturing of meat processing product using Vegetable Lactobacillus |
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