甲醇蛋白联产木聚糖酶发酵培养基优化
Optimization of fermentation culture medium for methanol protein co-production xylanase
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摘要: 为将实验室诱变选育出的高产甲醇蛋白毕赤酵母菌高效表达木聚糖酶，在100 L发酵罐中，对影响木聚糖酶发酵水平的培养基组分及其质量浓度等因素进行考查，初步确定培养基配方，并设计正交试验优化发酵培养基.结果表明：最优发酵培养基配方为H3PO4质量浓度25 g/L、K2HPO4质量浓度0.5 g/L、NH4Cl质量浓度0.30 g/L、CaSO4· 2H2O质量浓度0.25 g/L、NaCl质量浓度0.6 g/L、K2SO4质量浓度3.0 g/L、MgSO4· 7H2O质量浓度2.2 g/L；在最优培养基配方下，菌体湿重达到409~417 g/L，木聚糖酶活力达到40 915~41 428 U/mL.Abstract: In order to produce the xylanase efficiently, the effects of the culture medium composition and content in a 100 L fermentation tank on the xylanase fermentation were investigated with the high-yield methanol protein strain of Pichia pastoris selected by mutagenesis in our lab. The ratio of the medium was determined. Then, the orthogonal experimental scheme was designed to optimize the ratio of the medium. The result showed that the optimal components of fermentation medium were mass concentration of phosphoric acid of 25 g/L, dipotassium hydrogen phosphate of 0. 5 g/L, ammonium chloride of 0. 3 g/L, calcium sulfate dihydrate of 0. 25 g/L, sodium chloride of 0. 6 g/L, potassium sulfate of 3. 0 g/L, and magnesium sulfate heptahydrate of 2. 2 g/L; under the optimal medium formula, the wet weight of fermentation reached 409~417 g/L, and the xylanase activity reached 40 915~41 428 U/mL.
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Key words:
- methanol protein /
- xylanase /
- Pichia pastoris /
- fermentation culture medium
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[1]
李双娟. 甲醇蛋白的生产及发展前景[J]. 中氮肥,2006(3):4.
-
[2]
谢军. 甲醇蛋白的营养价值及社会效益[J]. 煤炭与化工,2015,38(7):41.
-
[3]
谷小虎,李晓辉,杨旭,等. 甲醇蛋白的技术与市场[J]. 煤化工,2011,156(5):5.
-
[4]
徐兆瑜. 甲醇产业的发展和工艺技术新进展[J]. 精细化工原料及中间体,2007(6):23.
-
[5]
郭建华. 甲醇蛋白的技术经济分析与发展前景[J]. 化工技术经济,2003,21(6):25.
-
[6]
苟万晓,范延超,吕昂,等. 黑曲霉ZF-34脂肪酶基因克隆及其在毕赤酵母中的表达[J]. 河南工业大学学报(自然科学版),2019,40(5):59.
-
[7]
曹淼,胡全伟. 浅谈甲醇蛋白的开发和应用前景[J]. 化工管理,2016(20):65.
-
[8]
LI X,HE X Y,LI Z L,et al. Combined strategies for improving the production of recombinant Rhizopus oryzae lipase in Pichia pastoris[J]. Bioresources, 2013,8(2):2867.
-
[9]
毕荣宇,牟德华. 利用毕赤酵母生产单细胞蛋白的玉米粉-豆粕培养基优化[J]. 中国饲料, 2013(15):6.
-
[10]
WANG H L, LI J H, LIU L, et al. Increased production of alkaline polygalacturonate lyase in the recombinant Pichia pastoris by controlling cell concentration during continuous culture[J]. Bioresource Technology, 2012,124:338.
-
[11]
杜少平,胡海艳,甘祥武,等. 重组毕赤酵母产β-甘露聚糖酶的高密度发酵研究[J]. 轻工学报,2020,35(4):1.
-
[12]
魏涛,赵彩梦,郏未未,等. 果糖基转移酶AoFT在毕赤酵母中的表达与纯化及其酶学性质研究[J]. 轻工学报,2020,35(3):1.
-
[13]
李昆太,周佳,庞会忠,等. 热带假丝酵母SG-1产单细胞蛋白的发酵培养基优化[J]. 中国饲料,2013(3):15.
-
[14]
HERNÁNDEZ L,MENÉNDEZ C,PÉREZ E R, et al. Fructooligosaccharides production by Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase constitutively expressed to high levels in Pichia pastoris[J]. Journal of Biotechnology,2018,266:59.
-
[15]
沈伊娜,顾磊,张娟,等. 双碳源流加对重组毕赤酵母高效表达葡萄糖氧化酶的影响[J]. 生物工程学报,2013,29(7):927.
-
[16]
苟万晓,卫红伟,胡元森,等. 毕赤酵母发酵生产甲醇蛋白工艺条件的优化[J]. 中国酿造, 2014,33(7):78.
-
[17]
曹明,王杰,赵培,等. 毕赤酵母甲醇利用及甲醇蛋白发酵条件优化[J]. 河南科学,2015,33(5):731.
-
[18]
全国饲料工业标准化技术委员会. 饲料添加剂木聚糖酶活力的测定分光光度法:GB/T 23874-2009[S]. 北京:中国标准出版社, 2009.
-
[19]
罗健东,郑平香,陈修. 细胞内pH与细胞凋亡[J]. 生命科学,1997(5):218.
-
[20]
李淑彬,周仁超,钟英长. 金属离子对黄柄曲霉生长和抗真菌抗生素合成的影响[J]. 微生物学通报,2002(3):20.
-
[21]
陈菲. 一株蛋白酶产生菌的鉴定、酶性质研究和表达载体构建[D]. 郑州:河南工业大学, 2018.
-
[22]
邓宇,华兆哲,赵志军,等. 氮源对枯草芽孢杆菌 WSHDZ-01合成过氧化氢酶的影响[J]. 应用与环境生物学报, 2008(4):544.
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