烟草节杆菌肌酐酶发酵培养基优化

doi:10.13523/j.cb.20150813

中国生物工程杂志

2015, Vol. 35

Issue (8): 90-95 DOI: 10.13523/j.cb.20150813

技术与方法

烟草节杆菌肌酐酶发酵培养基优化

梁剑光, 贾佳红, 吴俊伟, 朱洁

常熟理工学院生物与食品工程学院常熟 215500

Optimization of Fermentation Medium for Creatininase Production by Arthrobacter nicotiana

LIANG Jiang-guang, JIA Jia-hong, WU Jun-wei, ZHU Jie

School of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, China

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摘要：

对烟草节杆菌发酵产酶培养基进行了优化。在烟草节杆菌的初始发酵培养基条件下,肌酐酶初始酶活仅为9.2U/g湿菌。首先,通过肌酸诱导、金属离子筛选实验发现肌酸和金属离子对烟草节杆菌产肌酐酶酶活有重要影响;然后再通过碳源和氮源优化,以玉米浆和酵母膏作为复合氮源,可溶性淀粉为碳源,肌酐酶产量可达到108.5 U/g湿菌;在以上基础上,最后通过两次正交试验优化初始发酵培养基,最优培养基组成为:肌酸 0.3%,玉米浆0.3%,可溶性淀粉0.4%,酵母膏0.7%,Fe²⁺ 0.003%,Mn²⁺ 0.006%,Mg²⁺ 0.005%,K₂HPO₄ 0.1%,KCl 0.5%。在优化后的发酵培养基条件下,烟草节杆菌肌酐酶酶活达到182.82 U/g湿菌,为初始发酵培养基酶活的19.87倍。

关键词： 烟草节杆菌; 肌酐酶; 培养基优化

Abstract:

The optimal culture medium of creatininase production on the Arthrobacter nicotiana were carried out. Firstly, using creatine as inductor and metal ions experiment, it was found that creatine and metal ions have a great influence upon the creatininase production. Secondly, single factor experiment showed the optimization carbon and nitrogen sources was soluble starch and corn steep liquor,but compound nitrogen source of yeast extract and corn steep liquor was much better(creatininase production reached 108.5 U/g wet mycelium). At last, creatininase production reached 182.5 U/g wet mycelium under the optimization of fermentation medium, which was about 19.87 times as high as the initial fermentation enzyme activity.

Key words: Arthrobacter nicotiana Creatininase Optimization of production medium

收稿日期: 2015-03-23 出版日期: 2015-08-25

ZTFLH:

Q93-335

通讯作者: 梁剑光 E-mail: liang4523@126.com

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引用本文:

梁剑光, 贾佳红, 吴俊伟, 朱洁. 烟草节杆菌肌酐酶发酵培养基优化[J]. 中国生物工程杂志, 2015, 35(8): 90-95.

LIANG Jiang-guang, JIA Jia-hong, WU Jun-wei, ZHU Jie . Optimization of Fermentation Medium for Creatininase Production by Arthrobacter nicotiana. China Biotechnology, 2015, 35(8): 90-95.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150813 或 https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I8/90

[1] 崔有宏, 罗侃, 郑强, 等. 肌酐测定工具酶产生菌烟草节杆菌02181的筛选及其产酶条件. 甘肃科学学报, 2005, 17(2): 49-53. Cui Y H, Luo K, Zheng Q, et al. Arthrobacter nicotianae 02181 which produces enzymes for diagnostic measurement of creatinine and its selected conditions for producing enzymes. Journa1 of GanSu Science, 2005, 17(2): 49-53.

[2] Yamamoto K, Oka M, Kikuchi T, et al. Cloning of the creatinine amidohydrolase gene from Pseudomonas sp PSJ. Biosci Biotech Biochem, 1995, 59(7): 1331-1332.

[3] Koyama Y, Yamamoto Otake, Suzuki M, et al. Cloning and expression of the sarcosine oxidase gene from Bacillussp S-19 in Escherichia coli. J Agric Biol Chem, 1991, 55(5): 1259-1263.

[4] 李萍,赵莹,余霆.肌酐在肾小球滤过功能损伤诊断中的价值的系统评价. 中国循证医学杂志,2004,4(11):752-758. Li P, Zhao Y,Yu T. Diagnostic value of creatinine for indicating glomerular filtration function injury: a systematic review. Chinese Journal of Evidence-based Medicine, 2004,4(11):752-758.

[5] 智强, 李淑慧, 高利宏, 等. 利用简并PCR克隆烟草节杆菌02181肌酸酶基因. 第三军医大学学报, 2007,29(12):1221-1223. Zhi Q,Li S H,Gao L H,et al. Gene cloning of creatinase from Arthrobacter nicotianae 02181 using degenerate PCR. Acta Academiae Medicinae Militaris Tertiae, 2007,29(12):1221-1223.

[6] 崔有宏, 罗侃, 郑强,等. 肌酐测定的酶学方法. 西北国防医学杂志, 2003, 24(2):129-131. Cui Y H, Luo K, Zheng Q, et al. Enzymatic method for the determination of creatinine. Medical Journal of National Defense Forces in Northwest China, 2003, 24(2):129-131.

[7] Paurekh A C, Cook S, Sims C, et al. A new method for the determination of serum creatinine based on reaction with 3,5-dinitrobenzoylchloride in an organic medium. Clin Chim Acta, 1976, 73(2): 221-231.

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