海洋低温葡萄糖氧化酶生产菌种的选育及发酵条件优化
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摘要
从大连黄海海域海泥中筛选得到一株高产葡萄糖氧化酶(GOD)的酵母菌,并对其进行菌种鉴定及发酵条件优化。经筛选,得到一株产低温GOD的酵母菌,编号为WYQ 23,其酶活为0.66 U/m L。通过形态学及分子生物学鉴定,确定该菌株为担子菌(Basidioascus sp.)。通过单因素试验优化,确定该菌株产低温GOD最优产酶条件为葡萄糖4%、蛋白胨0.3%、NaNO_30.3%、Mg SO_40.07%、KCl 0.03%、KH_2PO_40.3%、CaCO_30.6%,初始pH值6.0,发酵温度25℃,培养时间38 h。在此优化条件下,该菌株产低温GOD酶活力达到1.67 U/m L,是优化前的2.53倍。
A high-yield glucose oxidase(GOD) yeast was screened from marine mud in the Yellow Sea of Dalian, and then the strain was identified and the fermentation conditions were optimized. After screening, the yeast producing low temperature GOD was obtained and numbered as strain WYQ23, and the enzyme activity was 0.66 U/ml. The strain was identified as Basidioascus sp. by morphological and molecular biology. The optimum fermentation conditions for low temperature GOD production were optimized by single factor experiments as follows: glucose 4%, peptone 0.3%,NaNO_30.3%, Mg SO_40.07%, KCl 0.03%, KH_2 PO_40.3%, CaCO_30.6%, initial pH 6.0, fermentation temperature 25 ℃ and time 38 h. Under the optimum conditions, the low-temperature GOD activity of the strain reached 1.67 U/ml, which was 2.53 times that of before optimization.
A high-yield glucose oxidase(GOD) yeast was screened from marine mud in the Yellow Sea of Dalian, and then the strain was identified and the fermentation conditions were optimized. After screening, the yeast producing low temperature GOD was obtained and numbered as strain WYQ23, and the enzyme activity was 0.66 U/ml. The strain was identified as Basidioascus sp. by morphological and molecular biology. The optimum fermentation conditions for low temperature GOD production were optimized by single factor experiments as follows: glucose 4%, peptone 0.3%,NaNO_30.3%, Mg SO_40.07%, KCl 0.03%, KH_2 PO_40.3%, CaCO_30.6%, initial pH 6.0, fermentation temperature 25 ℃ and time 38 h. Under the optimum conditions, the low-temperature GOD activity of the strain reached 1.67 U/ml, which was 2.53 times that of before optimization.
引文
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[24]石淑钰,张庆芳,迟乃玉,等.一株海洋低温葡萄糖氧化酶菌株的筛选、鉴定及部分酶学性质[J].微生物学通报,2014,41(5):832-838.
[25]王晓辉,王贵鹏,张庆芳,等.一株抗灰霉病解淀粉芽孢杆菌的筛选鉴定及抑菌蛋白的分离[J].吉林农业科学,2015,40(1):64-67.
[2]叶日英,徐德峰,孙力军,等.海洋源高产葡萄糖氧化酶细菌的筛选和主要酶学性质[J].浙江农业学报,2018,30(4):672-678.
[3]李蓉,张庆芳,迟乃玉.微生物葡萄糖氧化酶的研究进展[J].中国酿造,2018,37(3):1-5.
[4]SINGH O V.Mutagenesis and analysis of mold Aspergillus niger for extracellular glucose oxidase production using sugarcane molasses[J].Appl Biochem Biotech,2006,135(1):43-57.
[5]BELYAD F,KARKHANEI A A,RAHEB J.Expression,characterization and one step purification of heterologous glucose oxidase gene from Aspergillus niger ATCC 9029 in Pichia pastoris[J].EuPA Open Proteom,2018,19:1-5.
[6]邵彩梅,刘燕,王玉璘,等.葡萄糖氧化酶对丹系母猪围产期便秘、粪便菌群及初乳品质的影响[J].饲料工业,2018,39(21):39-43.
[7]惠明,董贞,田青,等.碳纤维共固定葡萄糖氧化酶-过氧化氢酶的研究[J].中国食品学报,2018,18(3):134-140.
[8]龚金秋,刘志强,田雯,等.葡萄糖氧化酶的研究进展及其在猪生产中的应用[J].中兽医医药杂志,2018,37(5):34-38.
[9]王冰.葡萄糖氧化酶在猪饲料中的研究进展[J].饲料博览,2018(1):21-24.
[10]胡亚奇.葡萄糖氧化酶在鸡生产上的应用[J].中兽医学杂志,2018(1):76.
[11]张同燕.葡萄糖氧化酶在养殖业中的应用研究进展[J].当代畜牧,2018(3):16-17.
[12]黄晓月,莫日坚,徐德峰,等.海洋源Bacillus cereus CAMT2377产葡萄糖氧化酶过程分析及优化[J].中国生物制品学杂志,2018,31(11):1268-1273.
[13]郭云瑕,孙钰婷,辛颖,等.葡萄糖氧化酶产生菌的紫外诱变及发酵条件优化[J].食品科技,2016,41(5):2-6.
[14]HAQ U,NAWAZ I,MUKHTAR A,et al.Optimization of inoculum volume,fermentation medium and aeration rate for the production of glucose oxidase by UV mutant strain of Aspergillus niger an-14[J].Pakistan J Bot,2015,47(SI):329-332.
[15]范新蕾,肖成建,顾秋亚,等.响应面法优化黑曲霉产葡萄糖氧化酶发酵条件[J].食品与生物技术学报,2014,33(10):1096-1100.
[16]郭贺楠,杨勇智,董冰,等.葡萄糖氧化酶研究进展[J].中国畜牧杂志,2018,54(4):10-14.
[17]高庆华,董聪,王玥,等.共表达分子伴侣PDI和Ero1对葡萄糖氧化酶在毕赤酵母中表达的影响[J].生物技术通报,2018,34(7):174-179.
[18]顾磊.Aspergillus niger葡萄糖氧化酶的异源分泌表达、分子改造和发酵生产[D].无锡:江南大学,2014.
[19]闻一凡,顾磊,张娟,等.定点突变提高毕赤酵母产葡萄糖氧化酶的氧化稳定性[J].食品与生物技术学报,2016,35(12):1260-1267.
[20]KHATTAB A A,BAZARAA W A.Screening,mutagenesis and protoplast fusion of Aspergillus niger for the enhancement of extracellular glucose oxidase production[J].J Ind Microb Biot,2005,32(7):289-294.
[21]范新蕾,肖成建,顾秋亚,等.ARTP诱变选育葡萄糖氧化酶高产菌株及发酵条件优化[J].工业微生物,2015,45(1):15-19.
[22]王周圆,别小妹,吕凤霞,等.产黄青霉A4产胞外葡萄糖氧化酶发酵工艺优化[J].食品工业科技,2015,36(16):217-221.
[23]朱洪菊.葡萄糖氧化酶产生菌的诱变育种及条件优化[D].济南:山东师范大学,2012.
[24]石淑钰,张庆芳,迟乃玉,等.一株海洋低温葡萄糖氧化酶菌株的筛选、鉴定及部分酶学性质[J].微生物学通报,2014,41(5):832-838.
[25]王晓辉,王贵鹏,张庆芳,等.一株抗灰霉病解淀粉芽孢杆菌的筛选鉴定及抑菌蛋白的分离[J].吉林农业科学,2015,40(1):64-67.