Abstract: It has been known that the furfural and acetic acid in the liquid corn stalk hydrolysate could reduce yeast activity. The purpose of this study was to screen a furfural and acetic acid-resistant, high -yield ethanol Saccharomyces cerevisiae through mutation and domestication. A mutant library derived from Saccharomyces cerevisiae xp was acquired through atmospheric and room-temperature plasma (ARTP) mutagenesis. Subsequently, the mutants were grown in corn stalk hydrolysate liquid medium. After 25 generations of continuous domestication, a strain, named xp2, was obtained. This domesticated mutant had a maximum biomass DCW of 3.71 g/L, which was 19.68% higher than the wild type (3.10 g/L). It also had a short growth cycle, with 36 hours, which was 20 hours shorter than the wild type. The concentrations of the furfural and acetic acid in supernatant were 1.43 g/L and 1.21 g/L, which were 62.17% and 26.67% lower than those of the wild type (3.78 g/L and 1.65 g/L) respectively. These results indicated that the transformation capacity of the furfural and acetic acid had been significantly improved. The ethanol yield and average yield of xp2 were 38.7 g/L and 0.806 g/(L·h), which increased by 17.12% and 27.71% than that of the wild type. In conclusion, the screened strain in this study had high transformation capacity for furfural and acetic acid, and the screening method provided valuable reference for screening high performance Saccharomyces cerevisiae.
陈胜杰,高翔,袁戎宇. 常温常压等离子诱变结合玉米秸秆水解液驯化酿酒酵母生产生物乙醇[J]. 食品与发酵工业, 2020, 46(4): 167-171.
CHEN Shengjie,GAO Xiang,YUAN Rongyu. Screening of high-yield ethanol Saccharomyces cerevisiae by the combination of atmospheric and room temperature plasma with corn stalk hydrolysate[J]. Food and Fermentation Industries, 2020, 46(4): 167-171.
 忻胜兵.优质特性酿酒酵母的选育及直投式发酵菌剂研发[D]. 呼和浩特:内蒙古农业大学, 2018.  ZHU J, LI R, ZHANG Z, et al. Temporal and spatial distribution of crops straw and its comprehensive utilization mechanism in Shaanxi[J]. Transactions of the Chinese Society of Agricultural Engineering, 2013, 29(1):1-9.  舟丹. 龙力生物首家生产二代纤维素燃料乙醇[J]. 中外能源, 2018(11):59.  LEE J W, KIM J Y,JIANG H M, et al. Sequential dilute acid and alkali pretreatment of corn stover: Sugar recovery efficiency and structural characterization[J]. Bioresource Technology, 2015, 182:296-301.  SASANO Y, WATANABE D, UKIBE K, et al. Overexpression of the yeast transcription activator Msn2 confers furfural resistance and increases the initial fermentation rate in ethanol production[J]. Journal of Bioscience & Bioengineering, 2012, 113(4):451-455.  陈尚钘, 勇强, 徐勇, 等. 玉米秸秆稀酸预处理的研究[J]. 林产化学与工业, 2009, 29(2):27-32.  朱艳, 王奇, 原帅, 等. 响应曲面法优化稀硫酸预处理玉米秸秆[J]. 西安工程大学学报, 2015(3): 301-306.  李小娟, 黄瑞, 张超,等. 定向驯化筛选耐毒酵母[J]. 食品工业科技, 2014, 35(7):163-167.  ZHANG X, ZHANG C, ZHOU Q Q, et al. Quantitative evaluation of DNA damage and mutation rate by atmospheric and room-temperature plasma (ARTP) and conventional mutagenesis[J]. Applied Microbiology & Biotechnology, 2015, 99(13):5 639-5 642.  LI H P, WANG Z B, GE N, et al. Studies on the Physical Characteristics of the radio-frequency atmospheric-pressure glow discharge plasmas for the genome mutation of methylosinus trichosporium[J]. IEEE Transactions on Plasma Science, 2012, 40(11):2 853-2 860.  JIN M, GUNAWAN C, UPPUGUNDLA N, et al. A novel integrated biological process for cellulosic ethanol production featuring high ethanol productivity, enzyme recycling and yeast cells reuse[J]. Energy & Environmental Science, 2012, 5(5):7 168-7 171.  张素, 肖洋, 吉玉玉, 等. 高效液相色谱法测定黑曲霉xj菌丝体中糠醛的含量[J]. 中国酿造, 2018, 37(5):181-183.  陈胜杰, 刘辉, 谢希贤, 等. 敲除iclR基因对大肠杆菌发酵L-色氨酸的影响[J]. 天津科技大学学报, 2016, 31(3):25-30.  陈牧, 连之娜, 徐勇, 等. 硫酸水解-高效液相色谱法定量测定低聚木糖[J]. 生物质化学工程, 2010, 44(6):14-17.  林贝, 李健秀, 刘雪凌,等. 紫外诱变结合驯化提高酿酒酵母对抑制物耐受性[J]. 生物技术, 2018, 28(1): 85-88.  ZHANG M, WANG F, SU R, et al. Ethanol production from high dry matter corncob using fed-batch simultaneous saccharification and fermentation after combined pretreatment [J]. Bioresource Technology, 2010, 101(13):4 959-4 964.  徐亚男.非酿酒酵母菌的筛选及产酶研究[D]. 石河子:石河子大学, 2016.  ARKELL A, KRAWCZYK H, JNSSON A S. Influence of heat pretreatment on ultrafiltration of a solution containing hemicelluloses extracted from wheat bran[J]. Separation & Purification Technology, 2013, 119(46):46-50.  SINGH J, SUHAG M, DHAKA A. Augmented digestion of lignocellulose by steam explosion, acid and alkaline pretreatment methods: A review[J]. Carbohydr Polym, 2015, 117:624-631.  YIN J, HAO L, TU W Y, et al. Enzymatic hydrolysis enhancement of corn lignocellulose by supercritical CO2 combined with ultrasound pretreatment[J]. Chinese Journal of Catalysis, 2014, 35(5):763-769.