Optimization of Wickerhamomyces anomalus Fermentation Conditions for Pectinase Production Based on Wet Processing of Arabica Coffee in Yunnan Province  

基于云南小粒咖啡湿法加工的异常威克汉姆酵母产果胶酶发酵条件优化

作　　者：ZHANG Shu-zhu MENG Jin XUE Xiao-ran KONG De-ting TIE Li-ping HUANG Jia-xiong QIN Shi-wen HE Fei-fei SUO Yu-kai 张树竹;蒙锦;薛笑然;孔德婷;铁丽萍;黄家雄;秦世雯;何飞飞;索玉凯(云南大学资源植物研究院,农学院,云南昆明650000;云南民族大学民族医药学院民族药资源化学国家民委-教育部重点实验室,云南昆明650000;云南民族大学民族药内生菌天然产物合成生物学国家民委重点实验室,云南昆明650000;云南省农科院热带和亚热带经济作物研究所,云南普洱665000)

机构地区：[1]School of Agriculture,Yunnan University,Kunming 650000,PRC [2]Key Laboratory of Chemistry in Ethnic Medicinal Resources,State Ethnic Affairs Commission and Ministry of Education,Yunnan Minzu University,Kunming 650000,PRC [3]Key Laboratory of Synthetic Biology of Ethnic Medicinal Endophyte-derived Natural Products,State Ethnic Affairs Commission,Yunnan Minzu University,Kunming 650000,PRC [4]Institute of Tropical and Subtropical Economic Crops,Yunnan Academy of Agricultural Sciences,Pu’er 665000,PRC

出　　处：《Agricultural Science & Technology》2022年第3期30-39,共10页农业科学与技术（英文版）

基　　金：云南省科技计划项目(202202AE 090002);云南大学新一轮“双一流”建设项目(CZ22634404);云南大学研究生科研创新项目(2021Y429,2021Y251)。

摘　　要：In order to improve the pectin-degrading efficiency in wet processing of Arabica coffee in Yunnan, Box-Behnken design and single factor experiment were used to optimize the fermentation conditions of five pectinolytic Wickerhamomyces anomalus strains from the fermentation broth of Arabica coffee in Baoshan, Yunnan during wet processing with pectase activity as an indicator. The results showed that the five strains all synthesized pectin lyase(PL), polygalacturonase(PG), and pectin methylesterase(PM).Among them, strain CAP5 had strong ability to produce PG and PL,while strain CAP4 secreted a large amount of PM. Under optimized conditions, the activity of PG, PL, and PM of the five strains came in at 250.17~411.20 U/mL, 12.98~16.55 U/mL, and 208.52~322.83 U/mL,respectively. The four factors of nitrogen source concentration,fermentation time, Mn2+ concentration, and pH value were optimized and the optimal pectinase-producing fermentation conditions for five strains were as follows: peptone 2.2 g/L, fermentation time 30 h, Mn2+ 1.5 mmol/L, and pH 4.3. After fermentation under the optimized conditions, the maximum PG activity of CAP5 amounted to 411.20 U/mL, 114.03% higher than that before optimization.Meanwhile, the PG activity of strains CAP3, CAP4, CAP8, and CAP10 increased by 86.74%, 114.55%, 65.79%, and 66.07%,respectively, and the activity of PL and PM of the five strains rose 150.35%~218.56% and 341.07%~418.52%, respectively. These findings suggested that W. anomalus strains could be used as coffee starter and had great potential for the lysis of pectin.为提高异常威克汉姆酵母(Wickerh-amomyces anomalus)在云南小粒咖啡湿法加工中的脱胶效率,以果胶酶活性为指标,通过Box-Behnken设计和单因素试验对云南保山小粒咖啡湿法发酵液中获得的5株产果胶酶W. anomalus菌株的发酵条件进行优化。结果表明:5株酵母均可产生果胶裂解酶(PL)、多聚半乳糖醛酸酶(PG)和果胶甲酯酶(PM),活性分别达到250.17~411.20 U/mL、12.98~16.55 U/mL和208.52~322.83 U/mL。CAP5菌株具有较高的PG和PL酶活,而CAP4菌株具有较高的PM酶活。通过对氮源浓度、发酵时间、Mn2+浓度和pH值进行优化,获得最优产果胶酶发酵条件:蛋白胨浓度2.2 g/L,发酵时间30 h,Mn浓度1.5 mmol/L,pH 4.3。优化后,CAP5菌株最大PG活性可达411.20 U/mL,较优化前增加114.03%。CAP3、CAP4、CAP8和CAP10菌株的PG活性较优化前分别增加86.74%、114.55%、65.79%和66.07%。5株酵母的PL和PM活力比优化前分别提高150.35%~218.56%和341.07%~418.52%。以上结果说明W. anomalus菌株可作为咖啡发酵剂,具有较好的脱胶潜力。

关 键 词：Wickerhamomyces anomalus COFFEE Wet processing PECTINASE Box–Behnken design 

分 类 号：TS273[轻工技术与工程—农产品加工及贮藏工程]