Screening of Xylanase-producing Bacillus sp. and Optimization of Fermentation Process
-
摘要: 利用木聚糖酶选择培养基, 从120株标准芽胞杆菌中筛选出37株能够形成明显透明圈的芽胞杆菌。经产酶发酵液培养基复筛, 得到1株木聚糖酶活性较高的菌株FJAT-14260 (沙福芽胞杆菌Bacillus safensis) 。采用单因素实验设计和正交实验设计进行优化, 结果表明, 在培养基优化中对产酶影响最大的因素是底物浓度;在发酵条件优化中对产酶影响最大的因素是培养温度。优化得到FJAT-14260的最佳产酶发酵条件为:碳源 (酵母粉) 7g·L-1, 氮源 (蛋白胨) 9g·L-1, 底物 (木聚糖) 浓度10g·L-1, 培养温度30℃, 初始pH值8, 装液量30mL/250mL。在此条件下FJAT-14260在发酵32h时酶活力达到113 585.78U·mL-1, 比优化前提高了4.62倍。Abstract: Selective medium for xylanase was used in screening the xylanase producing Bacillus strains.Thirtyseven of the 120 Bacillus strains tested were found to form transparent circles on the plates.After a secondary screening with the enzyme fermentation medium, B.safensis FJAT-14260 was determined to be the most desirable strain for the xylanase production.A single factor and an orthogonal experiment were used to optimize the fermentation conditions.The results showed the substrate concentration to be the most important factor relating to the medium, and temperature, the most critical condition for the fermentation process.In all, the best medium for the enzyme production consisted of carbon source (yeast extract) at 7g·L-1, nitrogen source (peptone) at 9g·L-1, and substrate (xylan) at 10g·L-1.And, the optimal fermentation temperature was 30℃, initial pH at 8.0, and liquid volume at 30mL/250 mL.Under these conditions, after allowing FJAT-14260 to ferment for 32 h, the xylanase activity could reach as high as 113 585.78U·mL-1, which was 4.62 times of what was without the optimization.
-
Key words:
- Bacillus sp /
- xylanase /
- fermentation conditions
-
[1] SONG Y, LEE Y G, ChOI I S, et al.Heterologous expression of endo-1, 4-β-xylanase A from Schizophyllum communein Pichia pastoris and functional characterization of the recombinant enzyme[J].Enzyme and Microbial Technology, 2013, 52 (3) :170-176. [2] PRADE R A.Xylanases:from biology to biotechnology[J].Biotech GenEng Rev, 1995, 13:100-131. [3] COLLINS T, GERDAY C, FELLER G.Xylanases, xylanase families and extremophilic xylanases[J].FEMS Microbiol Rev, 2005, 29:3-23. [4] HRISTOV A N, MCALLISTER T A, CHENG K J.Intraruminal supplementation with increasing levels of exogenous polysaccharide-degrading enzymes:Effects on nutrient digestion in cattle fed a barley grain diet[J].J Anim Sci, 2000, 78 (2) :477–487. [5] 怀文辉, 何秀萍, 郭文洁, 等.微生物木聚糖降解醉研究进展及应用前景[J].微生物学报通报, 2000, (2) :137-139. [6] COURSON C, UDRON L, PETIT C, et al.Hydrogen production from biomass gasification on nickel catalyst:test for dry reforming of methane[J].Catalysis Today, 2002, 76 (1) :75-86. [7] 王秋影, 廖美德.木聚糖酶的研究进展[J].饲料博览, 2013, (6) :8-12. [8] TIMOTHY S, MAGNUSON, DON L CRAWFORD.Purification and characterization of an alkaline xylanase from streptomyces viridosporus T7A[J].Enzyme and Microbial Technology, 1997, 21:161. [9] 毕瑞明, 孙迅, 任少亭, 等.黑曲霉产木聚糖酶发酵条件的研究[J].工业微生物, 2000, 30 (1) :53. [10] 高庆义, 王效忠, 毕瑞明, 等.链霉菌发酵麦草产木聚糖酶的实验研究[J].工业微生物, 2001, 31 (3) :36. [11] 陆健.木聚糖酶的产生、性质和应用[J].酿酒, 2001, 28 (6) :30-34. [12] 包怡红, 李雪龙, 杨传平.类芽胞杆菌木聚糖酶产生菌株的筛选及其产酶条件优化[J].东北林业大学学报, 2008, 36 (9) :70-73. [13] 崔月明, 樊妙姬, 栾桂龙, 等.枯草芽胞杆菌XY1905木聚糖酶酶学性质的初步研究[J].饲料工业, 2005, 26 (6) :21-23. [14] 许正宏, 白云玲, 孙微, 等.细菌木聚糖酶高产菌的选育及产酶条件[J].微生物学报, 2000, 40 (4) :440-443. [15] BAILEY M J, BIELY P, POUTANEN K J.Interlabtoratory testing of methods for assay of xylanase[J].Biotechnol, 1992, 23:257-270. [16] 张彬, 林炜, 尚卓, 等.芽胞杆菌B50产高活性木聚糖酶的发酵条件研究[J].食品科技, 2007, (12) :39-42. [17] 李蕤, 程海燕, 张洁, 等.产碱性木聚糖酶芽胞杆菌HSI的筛选及发酵条件研究[J].工业微生物, 2002, 32 (2) :30-33. [18] 解复红.木聚糖酶产生菌的筛选和酶学性质研究[D].昆明:云南大学, 2003. [19] 管斌, 丁友昉, 谢来苏, 等.还原糖测定方法的规范[J].无锡轻工业大学学报, 1999, (3) :74-79. [20] 葛兰芳.木聚糖酶产生菌的筛选与发酵条件的初步优化.[D].成都:四川师范大学, 2008. [21] 孙振涛, 刘建军, 赵祥颖, 等.一株产木聚糖酶菌株的分离、鉴定及其酶学特性研究[J].生物技术, 2007, 17 (4) :74-77. [22] 单春乔, 曲恩玉, 江庆国, 等.产木聚糖酶、纤维素酶芽胞杆菌的分离、筛选及鉴定[J].黑龙江畜牧兽医, 2012, (2) :93-95.
点击查看大图
计量
- 文章访问数: 162
- HTML全文浏览量: 53
- PDF下载量: 2
- 被引次数: 0