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Directly mining a fungal thermostable alpha-amylase from Chinese Nong-flavor liquor starter

文献类型：期刊论文

作者Yi, Zhuolin2,3,4; Fang, Yang2,4; He, Kaize2,4; Liu, Dayu3; Luo, Huibo5; Zhao, Dong1; He, Hui6; Jin, Yanling2,4; Zhao, Hai2,3,4
刊名MICROBIAL CELL FACTORIES
出版日期2019-02-22
卷号17页码:30
关键词Chinese Nong-flavor liquor starter Thermostable Fungal alpha-amylase Starch degradation Solid-state simultaneous saccharification and fermentation
ISSN号1475-2859
DOI10.1186/s12934-018-0878-y
产权排序2
文献子类Article
英文摘要Background: Chinese Nong-flavor (NF) liquor is continuously and stably produced by solid-state fermentation technology for 1000 years, resulting in enrichment of special microbial community and enzymes system in its starter. Based on traditional culture-dependent methods, these functional enzymes are hardly obtained. According to our previous metatranscriptomic analysis, which identifies plenty of thermostable carbohydrate-active enzymes in NF liquor starter, the aim of this study is to provide a direct and efficient way to mine these thermostable enzymes. Results: In present study, an alpha-amylase (NFAmy13A) gene, which showed the highest expression level of enzymes in starch degradation at high temperature stage (62 degrees C), was directly obtained by functional metatran-scriptomics from Chinese Nong-flavor liquor starter and expressed in Pichia pastoris. NFAmy13A had a typical signal peptide and shared the highest sequence identity of 64% with alpha-amylase from Aspergillus niger. The recombinant enzyme of NFAmy13A showed an optimal pH at 5.0-5.5 and optimal temperature at 60 degrees C. NFAmy13A was activated and stabilized by Ca2+, and its half-lives at 60 and 70 degrees C were improved significantly from 1.5 and 0.4 h to 16 and 0.7 h, respectively, in the presence of 10 mM CaCl2. Meanwhile, Hg2+, Co2+ and SDS largely inhibited its activity. NFAmy13A showed the maximum activity on amylopectin, followed by various starches, amylose, glycogen, and pullulan, and its specificity activity on amylopectin was 200.4 U/mg. Moreover, this alpha-amylase efficiently hydrolyzed starches (from corn, wheat, and potato) at high concentrations up to 15 mg/ml. Conclusions: This study provides a direct way to mine active enzymes from man-made environment of NF liquor starter, by which a fungal thermostable alpha-amylase (NFAmy13A) is successfully obtained. The good characteristics of NFAmy13A in degrading starch at high temperature are consistent with its pivotal role in solid-state fermentation of NF liquor brewing. This work would stimulate mining more enzymes from NF liquor starter and studying their potentially synergistic roles in NF liquor brewing, thus paving the way toward the optimization of liquor production and improvement of liquor quality in future.
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WOS关键词BIOCHEMICAL-CHARACTERIZATION ; THERMOMYCES-LANUGINOSUS ; PHYSICOCHEMICAL PROPERTIES ; STARCH HYDROLYSIS ; PURIFICATION ; ENZYMES ; STRAIN ; FERMENTATION ; GLUCOAMYLASE ; EXPRESSION
WOS研究方向Biotechnology & Applied Microbiology
语种英语
WOS记录号WOS:000426386000001
出版者BIOMED CENTRAL LTD
源URL[http://210.75.237.14/handle/351003/30328]  
专题环境治理与食品安全领域_应用与环境微生物研究
环境治理与食品安全领域_环境微生物四川省重点实验室
环境治理与食品安全领域_中国科学院环境与应用微生物重点实验室
作者单位1.Wuliangye Grp, Yibin, Peoples R China;
2.Chinese Acad Sci, Chengdu Inst Biol, Environm Microbiol Key Lab Sichuan Prov, 9 Sect 4,Renmin Nan Rd, Chengdu 610041, Sichuan, Peoples R China;
3.Chengdu Univ, Coll Pharm & Biol Engn, Meat Proc Applicat Key Lab Sichuan Prov, Chengdu, Sichuan, Peoples R China;
4.Chinese Acad Sci, Key Lab Environm & Appl Microbiol, Chengdu, Sichuan, Peoples R China;
5.Sichuan Univ Sci & Engn, Bioengn Coll, Liquor Making Biotechnol & Applicat Key Lab Sichu, Zigong, Peoples R China;
6.Moutai Coll, Dept Liquor Making Engn, Renhuai, Peoples R China
推荐引用方式
GB/T 7714		 Yi, Zhuolin,Fang, Yang,He, Kaize,et al. Directly mining a fungal thermostable alpha-amylase from Chinese Nong-flavor liquor starter[J]. MICROBIAL CELL FACTORIES,2019,17:30.
APA Yi, Zhuolin.,Fang, Yang.,He, Kaize.,Liu, Dayu.,Luo, Huibo.,...&Zhao, Hai.(2019).Directly mining a fungal thermostable alpha-amylase from Chinese Nong-flavor liquor starter.MICROBIAL CELL FACTORIES,17,30.
MLA Yi, Zhuolin,et al."Directly mining a fungal thermostable alpha-amylase from Chinese Nong-flavor liquor starter".MICROBIAL CELL FACTORIES 17(2019):30.

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来源：成都生物研究所

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