高产右旋糖苷酶微生物的筛选及产酶条件的优化
Screening of high yield dextran microorganisms and optimization of enzyme production conditions
-
摘要: 使用营养缺陷型培养基、运用透明圈法从甘蔗土壤中筛得高产右旋糖苷酶的微生物SGC-2,根据菌株的形态和16S rDNA基因序列对其进行分析鉴定,并对其发酵产酶条件进行优化.结果表明,SGC-2为巨大芽孢杆菌(Bacillus megatherium),该菌株的适宜产酶条件为:右旋糖酐T2000质量浓度20 g/L,牛肉膏质量浓度10 g/L,NaCl质量浓度4 g/L,MgSO4质量浓度0.4 g/L,接种时间36 h,接种量3%,发酵时间96 h.该条件下,右旋糖苷酶酶活可高达28.32 U/mL,比优化前提高了151%.该右旋糖苷酶产生菌为首次报道,这为筛选高产右旋糖苷酶微生物的研究提供了一定的理论基础.Abstract: The microorganism SGC-2 with high yield of dextran glycosidase was screened from sugarcane soil using nutrient-deficient medium and transparent circle method. The microorganism SGC-2 was identified according to its morphology and 16S rDNA gene sequence analysis. Moreover, the conditions of fermentation and enzyme production were further optimized. The results showed that the microorganism SGC-2 was Bacillus megatherium. The optimum conditions for enzyme production were as follows: dextran T2000 20 g/L, beef extract 10 g/L, NaCl 4 g/L, MgSO4 0.4 g/L, inoculation time 36 h, inoculation volume 3%, fermentation time 96 h. Under this condition, the activity of dextran anhydrase reached 28.32 U/mL, which increased by 151% when compared with that before optimization. The dextran anhydrase-producing strain was firstly reported, which provided a theoretical basis for screening high-yield dextran microorganisms.
-
Key words:
- Polymer dextran /
- dextran /
- Bacillus megatherium /
- fermentation optimization
-
-
[1]
张洪斌,吴定涛,黄丽君,等.一株产右旋糖酐酶青霉的分离及酶的纯化和性质[J].微生物学报,2011,51:495.
-
[2]
EGGLESTON G,MONGE A.Optimization of sugarcane factory application of commercial dextranases[J].Process Biochemistry,2005,40:1881.
-
[3]
蒋丹,仇元新,胡涛,等.口腔链球菌右旋糖酐酶分子结构和功能的研究进展[J].国际口腔医学杂志,2008,35:249.
-
[4]
KIM Y M,SEO M Y, KANG H K,et al.Construction of a fusion enzyme of dextransucrase and dextranase:Application for one-step synthesis of isomalto-oligosaccharides[J].Enzyme and Microbial Technology,2009,44:159.
-
[5]
ASLAN Y,TANRISEVEN A.Immobilization of Penicillium lilacinum dextranase to produce isomaltooligosaccharides from dextran[J].Biochemical Engineering Journal,2007,34:8.
-
[6]
朱慧霞,王雅洁,邓胜松,等.绳状青霉菌发酵产右旋糖酐酶的条件研究[J].食品科学,2010,31:288.
-
[7]
SUGIURA M,ITO A,OGISO T,et al.Studies on dextranase:Purification of dextranase from Penicillium funiculosum and its enzymatic properties[J].BBA-Enzymology,1973,309:357.
-
[8]
PLESZCZYNSKA M,ROGALSKI J,SZCZODRAK J,et al.Purification and some properties of an extracellular dextranase from Penicillium notatum[J].Mycological Research,1996,100:681.
-
[9]
BARRETT J F,BARRETT T A.Purification and partial characterization of the multicomponent dextranase complex of streptococcus sobrinus and cloning of the dextranase gene[J].Infection & Immunity,1987,55:792.
-
[10]
KOBAYASHI M,TAKAGI S,SHIOTA M,et al.An isomaltotriose-producing dextranase from Flavobacterium sp M-73:purification and properties[J].Agricultural & Biological Chemistry,1983,47:2585.
-
[11]
焦豫良,王淑军,吕明生,等.产右旋糖酐酶埃氏交替单胞菌的筛选、鉴定及酶活分析[J].食品工业科技,2013,34:171.
-
[12]
吴定涛.高产右旋糖酐酶菌株的筛选及酶学性质和酶的应用研究[D].合肥:合肥工业大学, 2012.
-
[13]
ABDEL-NABY M A,ISMAIL A M S,ABDEL-FATTAH A M,et al.Preparation and some properties of immobilized Penicillium funiculosum 258 dextranase[J].Process Biochemistry,1999,34(4):391.
-
[14]
李卫娟.可降解牙菌斑生物膜氧化节杆菌右旋糖苷酶的研究[D].无锡:江南大学,2013.
-
[15]
吕明生,王淑军,房耀维,等.一株产低温右旋糖苷酶海洋细菌的筛选和鉴定[J].海洋科学,2011,35:32.
-
[16]
孙晋武.右旋糖酐酶及其应用[J].中国生物工程杂志,1986(4):35.
-
[17]
ZOHRA R R,AMAN A,ZOHRA R R,et al.Dextranase:hyper production of dextran degrading enzyme from newly isolated strain of Bacillus licheniformis[J].Carbohydrate Polymers,2013,92:2149.
-
[18]
王德龙.海洋微生物右旋糖苷酶的发酵和纯化研究[D].南京:南京农业大学,2014.
-
[19]
KUWAHARA T,NORIMATSU I,NAKAYAMA H,et al.Genetic variation in 16S-23S rDNA internal transcribed spacer regions and the possible use of this genetic variation for molecular diagnosis of Bacteroides species[J].Microbiology & Immunology,2013,45:191.
-
[20]
布坎南, 吉本斯.伯杰细菌鉴定手册[M].8版.北京:科学出版社,1984.
-
[1]
计量
- PDF下载量: 3
- 文章访问数: 1374
- 引证文献数: 0