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响应面法优化微小毛霉固态发酵生产凝乳酶工艺研
究
李倬林 1 邵淑娟 1,2 李铁柱 1 孙悦 3 李玉秋 1 王景会 1 郑丽 1,4 杨贞耐 1,2,4*
(1. 吉林省农业科学院农产品加工研究中心, 长春 130033; 2. 吉林大学军需科技学院, 长春 130062;3.
吉林农业大学; ,长春 130024)
摘要:本文采用麦麸培养基进行固态发酵,以研究发酵条件对产酶活力的影响,在此基础上应用
Plackett-Burman、最陡爬坡实验和 Box-Behnken 实验设计,对微小毛霉的固态发酵培养基进化以达到最佳
产酶量。实验结果表明,最佳发酵条件为:发酵时间 5 天,发酵温度 30℃,固液比值 ,加样量为 4 份,
测得凝乳酶酶活为 828U/mL。并且当麸皮质量为 , (NH4)2SO4 质量为 , KH2PO4 质量为 g 时,
微小毛霉产凝乳酶活力达到 1200U/mL。较未优化前活力提高了 %。
关键词:微小毛霉,固态发酵,凝乳酶,响应面法
Optimization of solid state fermentation using response surface
methodology for milk-clotting enzyme production by Mucor pusillus
Li Zhuolin1 ,Shao Shujuan1,2,Li Tiezhu1,Sun Yue3,LiYuqiou3,Wang Jinghui1,
Zheng Li 1,4, Yang Zhennai1,2,4*
(1. Jilin Academy of Agricultural Sciences, Center of Agro-food Technology,Changchun,130033;2.
College of Quartermaster and Technology, Jilin University,Changchun, 130062;3. Jilin
Agricultural University, Changchun , 130033;4. College of Biological and Agricultural
Engineering,Jilin University,Changchun,130024)
Abstract: Using Plackett-Burman bined with the path of steepest ascent method and Box-Behnken
design, the optimum ation properies including fermentation time 5d,fermentation temperature 30℃,
solid-to-liquid ratio and application with 4 samples were determined. Under such conditions, with increasing
the amount of bran, (NH4)2SO4 and KH2PO4 in the culture to , and respectively, the highest
enzyme milk-clotting activity 1200 U/mL was achieved, % higher than the milk-clotting activity before
optimization.
Keywords:Mucor pusillus; solid-state fermentation; milk-clotting enzyme; Response Surface Methodology
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凝乳酶是干酪及凝乳酶干酪素生产的关键性酶, 其主要生物学功能为专一性裂解
Phe105-Metl06 之间的肽键,从而导致牛奶的凝结,因此它被广泛应用于干酪和干酪素生产[1]。传
统的牛凝乳酶生产方法是通过屠宰未断奶的小牛并在其第四胃中提取,生产量较低难以满足现代工业
化生产干酪的需要[2,3]。相比于动物源凝乳酶,