广东工业大学学报 ›› 2017, Vol. 34 ›› Issue (02): 23-27.doi: 10.12052/gdutxb.160090

• 综合研究 • 上一篇    下一篇

黑曲霉液体发酵制备蛋白酶的条件优化

李杏华, 黄梓浩, 张驰, 毕艳梅, 丁利君, 黄德美, 刘丹   

  1. 广东工业大学 轻工化工学院, 广东 广州 510006
  • 收稿日期:2016-06-30 出版日期:2017-03-09 发布日期:2017-03-09
  • 通信作者: 丁利君(1965-),女,教授,主要研究方向为食品化学、食品加工.E-mail:qhdlj@gdut.edu.cn E-mail:qhdlj@gdut.edu.cn
  • 作者简介:李杏华(1991-),女,硕士研究生,主要研究方向为天然产物提取及分析.
  • 基金资助:

    广东省教育部产学研合作重大专项(2013A090100009)

An Optimized Design for Protease Produced by Aspergillus Niger Liquid-state Fermentation

Li Xing-hua, Huang Zi-hao, Zhang Chi, Bi Yan-mei, Ding Li-jun, Huang De-mei, Liu Dan   

  1. School of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou 510006, China
  • Received:2016-06-30 Online:2017-03-09 Published:2017-03-09

摘要:

采用麸皮、玉米粉、豆粕粉3种天然营养物质作为培养基材料,通过研究其组分配比,确定最佳培养基,通过单因素(加水量、发酵温度、发酵时间、pH)实验和四因素三水平正交试验优化发酵条件.研究结果表明,最佳培养基组成是:麸皮4 g、豆粕粉1 g、硫酸铵4 g、磷酸氢二钾0.3 g、氯化钙0.5 g;最适发酵条件是pH值7,加水量125 mL,在45℃下发酵120 h.在此条件下,黑曲霉液体发酵所生产的蛋白酶酶活力达到269.68 U/mL.

关键词: 黑曲霉, 蛋白酶, 液体发酵

Abstract:

Different proportion and combination of bran, corn flour and soybean powder was applied to optimize the best composition of culture medium. Single factor experiments and four factors and three levels orthogonal test were used to improve the fermentation conditions. The result showed that the optimal composition of culture medium was 4 g bran, 1 g soybean powder, 4 g ammonium sulphate, 0.3 g dipotassium hydrogen phosphate and 0.5 g calcium chloride, the protease production was maximized under the conditions with an addition of water at 45℃ for 120 h with pH 7. The optimal fermentation conditions obtained were the initial pH of 7.0, 125 mL addition of water, cultivation temperature of 45℃, and cultivation time of 120 h. Under this condition, the best protease activity of 269.68 U/mL was obtained.

Key words: Aspergillus niger, protease, liquid state fermentation

中图分类号: 

  • TS201.3

[1] 张士伟, 黄建飞, 罗立新. 枯草芽孢杆菌产蛋白酶发酵培养基的优化[J]. 中国酿造, 2013, 32(2):20-24.ZHANG S W, HUANG J F, LUO L X. Optimization of fermentation medium for protease production by Bacillus subtilis[J]. China Brewing, 2013, 32(2):20-24.
[2] PRABHA M S, DIVAKAR K, PRIYA J D A, et al. Statistical analysis of production of protease and esterase by a newly isolated Lysinibacillus fusiformis AU01:purification and application of protease in sub-culturing cell lines[J]. Annals of Microbiology, 2014:1-14.
[3] 陈显玲. 蛋白酶产生菌的筛选及发酵条件的优化[J]. 广州化工, 2014, 42(7):67-70.CHEN X L. Optimization of screening of protease producing strain and the fermentation condition[J]. Guangzhou Chemical Industry, 2014, 42(7):67-70.
[4] HAN S J, PARK H, KIM S, et al. Enhanced production of protease by Pseudoalteromonas Arctica PAMC 21717 via statistical optimization of mineral components and FED-batch fermentation[J]. Preparative Biochemistry & Biotechnology, 2016, 46(4):328-335.
[5] KASANA R C. Proteases from psychrotrophs:an overview[J]. Critical Reviews in Microbiology, 2010, 36(2):134-145.
[6] MADHUMITHAH C G, KRITHIGA R, SUNDARAM S, et al. Utilization of vegetable wastes for production of protease by solid state fermentation using Aspergillus niger[J]. World Journal of Agricultural Sciences, 2011, 7(5):550-555.
[7] 李曹龙, 柴秀娟, 杨晶, 等. 响应面法优化产蛋白酶菌株的发酵条件[J]. 中国酿造, 2013, 32(6):89-100.LI C L, CHAI X J, YANG J, et al. Optimization for fermentation conditions of protease-producing strain by response surface analysis[J]. China Brewing, 2013, 32(6):89-100.
[8] 顾瑾麟, 张红发, 夏永军, 等. 响应面法优化嗜麦芽窄食单胞菌产蛋白酶发酵培养基[J]. 食品工业科技, 2013, 34(24):168-172.GU J L, ZHANG H F, XIA Y J, et al. Optimization of fermentation medium for protease production by Stenotrophomonas maltophilia using response surface methodology[J]. Science and Technology of Food Industry, 2013, 34(24):168-172.
[9] NIYONZIMA F N, MORE S. Detergent-compatible proteases:microbial production, properties, and stain removal analysis[J]. Preparative Biochemistry and Biotechnology, 2015, 45(3):233-258.
[10] ZAMBARE V P, NILEGAONKAR S S, KSHIRSAGAR P R, et al. Scale up production of protease using Pseudomonas aeruginosa MCM B-327 and its detergent compatibility[J]. Journal of Biochemical Technology, 2014, 5(2):698-707.
[11] 程德勇, 缪礼鸿. 高产蛋白酶活性的枯草芽孢杆菌筛选及鉴定[J]. 武汉轻工大学学报, 2014, 33(1):6-10.CHENG D Y, MIAO L H. Screening and identification of high yield protease producing Bacillus strains[J]. Journal of Wuhan Polytechnic University, 2014, 33(1):6-10.
[12] 刘颖, 张彬彬, 孙冰玉, 等. 枯草芽孢杆菌高产中性蛋白酶发酵条件的优化[J]. 食品科学, 2014, 35(13):166-170.LIU Y, ZHANG B B, SUN B Y, et al. Optimization of fermentation conditions for production of neutral protease by Bacillus subtilis 10075[J]. Food Science, 2014, 35(13):166-170.
[13] KUMAR S, SHARMA N S, SAHARAN M R, et al. Extracellular acid protease from Rhizopus Oryzae:purification and characterization[J]. Process Biochemistry, 2005, 40(5):1701-1705.
[14] NILEGAONKAR S S, ZAMBARE V P, KANEKAR P P, et al. Production and partial characterization of dehairing protease from Bacillus Cereus, MCM B-326[J]. Bioresource Technology, 2007, 98(6):1238-1245.
[15] 莫重文, 黄岗. 固态发酵法生产发酵豆粕的研究[J]. 中国油脂, 2007, 32(7):38-40.MO C W, HUANG G. Production of fermented soybean meal by solid state fermentation[J]. China Oils and Fats, 2007, 32(7):38-40.
[16] RAJMALWAR S, DABHOLKAR P S. Production of protease by Aspergillus sp. using solid-state fermentation[J]. African Journal of Biotechnology, 2009, 8(17):4197-4198.
[17] 李祖明, 李鸿玉, 荣瑞芬, 等. 碱性蛋白酶生产菌的育种及其液态发酵条件的研究[J]. 食品研究与开发, 2008, 29(5):19-23.LI Z M, LI H Y, RONG R F, et al. Studies on the Breeding of Alkaline Protease Producing Strain and its Submerged Fermentation Conditions[J]. Food Research and Development, 2008, 29(5):19-23.
[18] 刘红梅, 师广波, 马艳芳, 等. 液态发酵法生产花生抗氧化肽和中性蛋白酶的工艺优化[J]. 食品工业科技, 2014, 35(18):175-179.LIU H M, SHI G B, MA Y F, et al. Optimization of the production of peanut antioxidative peptides and Neutrase using liquid state fermentation[J]. Science and Technology of Food Industry, 2014, 35(18):175-179.
[19] 瞿叶辉,丁利君,彭文媛,等. 响应面优化弹性蛋白酶酶解罗非鱼下脚料的工艺研究[J]. 广东工业大学学报,2016,33(5):77-82.QU Y H, DING L J, PENG W Y, et al. Optimization of enzymolysis of tilapia waste with response surface methdology by elastase[J]. Journal of Guangdong University of Technology, 2016, 33(5):77-82.
[20] 中国国家标准化管理委员会. GB/T 23527-2009蛋白酶制剂[S]. 北京:中国标准出版社, 2009.

[1] 瞿叶辉, 丁利君, 彭文媛, 刘珊珊, 黄金, 成晓玲. 响应面优化弹性蛋白酶酶解罗非鱼下脚料的工艺研究[J]. 广东工业大学学报, 2016, 33(05): 77-82.
[2] 谭竹钧; 沈明才; 张峰; 胡华平; . 胰蛋白酶抑制剂的研究进展[J]. 广东工业大学学报, 2007, 24(03): 6-9.
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