CN1281732C - Production of agar-agar enzyme by Alteromonas sp.nov.SY 37-12 strain - Google Patents
Production of agar-agar enzyme by Alteromonas sp.nov.SY 37-12 strain Download PDFInfo
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- CN1281732C CN1281732C CNB2004100236561A CN200410023656A CN1281732C CN 1281732 C CN1281732 C CN 1281732C CN B2004100236561 A CNB2004100236561 A CN B2004100236561A CN 200410023656 A CN200410023656 A CN 200410023656A CN 1281732 C CN1281732 C CN 1281732C
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
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Abstract
The present invention relates to a new microorganism, particularly to a novel microorganism, namely Alteromonas sp. nov. SY37-12 capable of generating agar enzymes, and an agar enzyme generated by the novel microorganism, particularly to a new strain, namely the Alteromonas sp. nov. SY37-12 derived from nature, and a method for producing an agar enzyme by utilizing the new strain. In the present invention, the strain, namely the Alteromonas sp. nov. SY37-12, is separated and purified from sea areas of south China sea, has the characteristic of generation of agar enzymes, and has shapes and physiological and biochemical properties which are all different from those of the known strains; the strain determined as a new strain of alternative monas according to the ninth edition of Bergey's Manual has the characteristics of wide nutritional requirement, easy culture and short culture time, and the generated agar enzyme has very high activity; by using the strain of the present invention as a producing strain of the agar enzyme, the method has the characteristics of high activity and stability of the agar enzyme, short production periodicity, low cost and realization of industrial production; agarose fragments with different molecular weight and size can be generated when the enzyme acts on agars.
Description
Technical field
The present invention relates to a kind of new microorganism Alteromonas sp.nov.SY37-12 and the agarase of this bacterial strain generation.
Background technology
By known to inspection information, the literature search, produce the bacterial classification main source Yu Haiyang of agarase according to the inventor.Cytophage, Vibrio; Streptomyces, Alteromonas, Pseudoalteromonas, some bacterial strain among the Pseudomonas all can produce agarase.According to uncle Jie Shi handbook the 9th edition, the form of bacterial strain of the present invention and physiological and biochemical property all are different from known bacterium, are from the isolated novel species of nature (this bacterial classification difference sees Table 1 with the feature of other this genus bacterium).
Summary of the invention
The purpose of this invention is to provide a kind of method that derives from the novel species Alteromonas sp.nov.SY37-12 of marine alga and utilize this bacterial strain production agarase.
Separation and purification of the present invention goes out the novel species Alteromonas sp.nov.SY37-12 that a strain derives from marine alga, and it has the characteristic that produces agarase, and depositary institution is called for short CCTCC, and preserving number is M204009.This agarase acts on the agarose fragment that agar-agar can produce the different molecular weight size.
Characteristics of the present invention are:
Separate the Alteromonas sp.nov.SY37-12 bacterial strain of producing agarase first.Fig. 1 has shown the electromicroscopic photograph of this bacterial strain.This bacterial strain is shaft-like, Gram-negative bacteria, 0.5～0.6 μ m * 1.8～2.1 μ m.End is given birth to single flagellum, and accumulating poly hydroxybutyric acid salt particle is not as stock in the born of the same parents.This bacterial classification derives from ocean environment, and growth must need NaCl.Determine that it is alternately zygosaccharomyces according to uncle Jie Shi handbook the 9th edition.This bacterial strain 37 ℃ of cultivation 24h on the 2216E substratum form single bacterium colony, bacterium colony oyster white, circle, neat in edge.It is wide that this bacterial classification has the nutritional requirement scope, and the short characteristics of cultivation and incubation time particularly have high inulinase-producing activity easily, the 16h that in the 2216E substratum, ferments, and the vigor of enzyme promptly reaches 300U/ml in its fermented liquid.With the production bacterial strain of bacterial strain of the present invention as agarase, have product activity height, good stability, with short production cycle, characteristics that cost is low, can realize industrialized production.
After the separation and purification, utilize SDS-PAGE to measure the molecular weight of the agarase of this bacterial strain generation.Fig. 2 shows that the molecular weight of this agarase is 39Kda.It is different from the molecular weight of the agarase of having reported as shown in Table 2.
Below by embodiment the present invention is described:
Embodiment
From marine site, South Sea collected specimens, carry out primary dcreening operation by the size of selecting bacterium colony on the 2216E substratum, to produce transparent circle, sieve a strain bacterium that obtains having high yield agarase vigor again by measuring enzyme activity.This bacterial strain is accredited as a novel species that replaces in the zygosaccharomyces by morphologic observation and Physiology and biochemistry.(the composition of 2216E substratum: yeast extract paste 1g, peptone 5g, high ferric phosphate 0.1g, agar-agar 15g, NaCl 20g, H
2O 1000ml)
With bacterial classification with the activation of 2216E substratum after, the inoculum size with 1% inserts and is equipped with in the 250ml triangular flask of 100ml fermention medium, behind 35 ℃ of shake-flask culture 16h, nutrient solution is removed thalline with refrigerated centrifuge in centrifugal 10 minutes with 4000rpm, obtains fermenting enzyme liquid.The condition of enzyme activity determination: 1mL enzyme liquid adds in the 20mL0.5% agar-agar substrate, 35 ℃ of reaction 30min.Boiling water bath deactivation 5min is with the amount of DNS method mensuration reducing sugar.Under above-mentioned reaction conditions, 1mL enzyme liquid 1min produces 1 μ g reducing sugar as an enzyme activity unit, with the D-semi-lactosi as standard.
The molecular weight determination that this bacterial classification produces agarase adopts (NH
4)
2SO
4The zymoprotein that obtains of precipitation is dialysed in the damping fluid of 50mmol Tris-HCl pH7.5, after the drying.Through DEAE-Sepharose ion-exchange chromatography and Sephacryl S-100 gel filtration chromatography separation and purification zymoprotein.SDS-PAGE obtains a single band, and molecular weight is 39Kda.
| A.aur- antia | A.ci- trea | A.colw- elliana | A.denit- rificans | A.esp- ejiana | A.halo- planktis | A.ha- nedai | A.lurteo- violacea | A.macl- eodii | A.nigrif- aciens | A.r- ubra | A.un- dina | A.sp. nov | |
| Cell shape is straight curved to have 4 ℃ of 35 ℃ 40 ℃ nitrate reduction denitrification aerogenesis generations of sheath flagellum growth thing fluorescence to need organic factor generation amylase alginic acid enzyme chitinase utilize the water-soluble citraurin water-soluble black element of the water-soluble uranidin of the D-Glucose D-MANNOSE D-galactolipin D-Fructose sucrose maltose cellobiose melibiose lactose salicin D-Glucose hydrochlorate N-ethyl glucuronide acid ammonium butanedioic acid fumaric acid DL-LACTIC ACID salt DL-glycerate L-ketoglutaric acid citrate aconitate PEARLITOL 25C glycerine L-threonine TYR Pidolidone L-arginine putrescine water-soluble purpurin of water-soluble brown pigment water-soluble red colouring matter | + - - + - - - - - + + - - + + - + - d - - - - - + - - - - - - - - - + - + - - - - + - | + - - - d d - - - + + d + + - + - - - - - - - - - - - - - - - - - - - + - - | + - - - - + - - + + - - - - - - - - - - + - - + - - + - - - - - - - + - - - - - | + - + + - - - + - + + + + + - - - + - - - - - - - - + + + - - + - - - - | + - - - d - - - - + + + - + d + d + + d + + - - - - - - - - + + + - + + - d - - - - - - - | + - - - d - d - - d d - d + + d d + + - - - - - + + + - - - + + d - - + d d - - - - - - - | + - + + - - + - + + - - + d - d - - - - - - - d + - - - + - - - - - - d + - + d - - - - - | + d d d + - - - - + + - + d - d - + - - - - - + - - d - - - - - + d - d - - + + - - | + - - - + d - - - - + d - + - + + + + + + + + + d - - d + - - - d + - + d d - - - - - - - | + - - - - - - - - + - - - - + + - - + + - + - + + + - - + + - + - + + - - - - - - - + | + - - - + - - - - + + - + + - - - - - - - - - + - - - - - - - + - - - - | - + - d - - - - - + d - + + - - - + + - - - - + + - - - - - - - + - d - - - - - - - | + - + - + - - - - + + - + + + - - - - - - - - - - + + - - - - - - - - - |
Table 1 is the discriminating between the zygosaccharomyces kind alternately
| Bacterial classification | Enzyme system | Molecular weight KDa |
| Vibrio sp.Strain JT0107 Alteromonas sp.E-1 Bacillus cereus ASK202 Vibrio sp.PO-303 4 Bacillus sp.MK03 Pseudomonas atlantica Vibrio sp.AP-2 Alteromonas sp.strain C-1. Pseudomonas sp.PT-5 Pseudomonas sp.strain W7 Alteromonas sp.nov.SY 37-12 | A B C | 72 82 90 87.5 115 57 92 32 20 52 31 59 39 |
The comparison of table 2Alteromonas sp.nov.SY 37-12 strain enzyme-producing and other strain enzyme-producings
Claims (2)
1, a kind of agarase is produced bacterial strain SY 37-12 (Alteromonas sp.nov.), and its preserving number is CCTCC M204009.
2, the application of the described bacterial strain of claim 1 in agarase is produced.
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| CNB2004100236561A CN1281732C (en) | 2004-03-02 | 2004-03-02 | Production of agar-agar enzyme by Alteromonas sp.nov.SY 37-12 strain |
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| CNB2004100236561A CN1281732C (en) | 2004-03-02 | 2004-03-02 | Production of agar-agar enzyme by Alteromonas sp.nov.SY 37-12 strain |
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| CN1560226A CN1560226A (en) | 2005-01-05 |
| CN1281732C true CN1281732C (en) | 2006-10-25 |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100591755C (en) * | 2005-06-23 | 2010-02-24 | 国家***第三海洋研究所 | Anti AIDS 1 type virus compound preparation method |
| CN101608166B (en) * | 2008-05-15 | 2011-03-16 | 汕头大学 | Flavobacterium strain and application thereof in generating agarase |
| CN101451113B (en) * | 2008-11-06 | 2010-10-06 | 国家***第二海洋研究所 | Vibrio natriegens and method for producing agarase by using the same |
| CN101955895B (en) * | 2010-02-09 | 2012-05-23 | 浙江工业大学 | Roseobactersp.zjut and application thereof in preparation of agaro-oligosaccharide |
| CN103451119B (en) * | 2012-06-03 | 2015-06-17 | 中国海洋大学 | Alteromonas and method thereby for producing gel-type enteromorpha polysaccharide degrading enzyme by using Alteromonas |
| CN103352016B (en) * | 2013-06-27 | 2015-03-11 | 中国海洋大学 | Method for preparing biological fertilizer by utilizing Alteromonas colwelliana A321 to ferment enteromorpha |
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