一株棉酚降解菌株的筛选鉴定及对棉粕的脱毒效果

doi:10.6048/j.issn.1001-4330.2022.12.021

新疆农业科学 ›› 2022, Vol. 59 ›› Issue (12): 3057-3065.DOI: 10.6048/j.issn.1001-4330.2022.12.021

• 作物遗传育种·耕作栽培·种质资源 • 上一篇下一篇

一株棉酚降解菌株的筛选鉴定及对棉粕的脱毒效果

张莉(), 杨晓龙, 王海亮, 南珊珊, 马雪儿, 聂存喜, 张文举(), 陈程()

石河子大学动物科技学院,新疆石河子 832000

收稿日期:2022-02-20 出版日期:2022-12-20 发布日期:2023-01-30
通信作者: 张文举,陈程
作者简介:张莉(1997-),女,新疆人,硕士研究生,研究方向为饲料资源的开发与利用,(E-mail)1278166647@qq.com
基金资助:
国家自然科学基金“棉铃虫棉酚降解酶的高活性筛选及其酶学特性与降解机理”(31860660);石河子大学高层次人才科研启动资金专项“基于棉酚降解相关基因的热带假丝酵母遗传转化及应用项目”(RCZK2019)

Selection and Identification of a Cotton Phenol Degradation Strain and Its Detoxification Effect on Cotton Meal

ZHANG Li(), YANG Xiaolong, WANG Hailiang, NAN Shanshan, MA Xueer, NIE Cunxi, ZHANG Wenju(), CHEN Cheng()

College of Animal Science and Technology, Shihezi University, Shihezi 832000, China

Received:2022-02-20 Online:2022-12-20 Published:2023-01-30
Correspondence author: ZHANG Wenju, CHEN Cheng
Supported by:
National Natural Science Foundation of China(31860660);A Special Scientific Research Launch Fund for High-Level Talents of Shihezi University“Genetic Transformation and Application Project of Tropical Pseudosilk Yeast Based on Genes Related to Cotton-Phenol Degradation”(RCZK2019)

摘要/Abstract

摘要： 【目的】 分离筛选具有降解棉酚能力的菌株,研究其生物学特性及发酵性能及棉粕脱毒效果,为棉粕脱毒生产和应用提供参考数据。【方法】 采用新疆棉籽粕,以棉酚为唯一碳源分离筛选具有高效降解棉酚的菌株,研究其生物学特性,综合形态学和16SrDNA序列测定鉴定试验菌株,应用该菌株发酵棉粕,分析其棉粕脱毒效果。【结果】 菌株JD-1最适温度为37℃、且随着时间的延长,菌株的生长浓度逐渐升高,在22~24 h间趋于平稳。但随时间的延长JD-1菌株的pH逐渐降低,趋于平稳。在24 h内随盐浓度的升高,菌株生长逐渐缓慢到最后趋于平稳,且具有一定的耐盐性。而过酸或过碱都会抑制其生长。在最适条件下,JD-1发酵棉粕降棉棉酚降解率为40%,其在进化关系上与乳酸片球菌属(Pediococcus lactis)同源性高达99%,确定JD-1菌株为乳酸片球菌属。【结论】 本文以棉酚为唯一碳源,报道了一株具有具有较好降解棉酚能力的乳酸片球菌,对棉粕中棉酚的脱毒率达40%。

关键词: 游离棉酚; 菌株筛选; 棉酚脱毒; 生物学特性

Abstract:

【Objective】 The purpose of this experiment is to isolate and screen strains with the ability to degrade gossypol, and to explore its biological characteristics, fermentation performance and cotton meal detoxification effec in hope of providing reference data for the production and application of cotton meal detoxification.【Methods】 Xinjiang cottonseed meal and gossypol as the sole carbon source was used to isolate and screen strains with high-efficiency degradation of gossypol, and study their biological characteristics, comprehensive morphology and 16SrDNA sequence determination were applied to identify the test strains, and then this strain was employed to ferment cottonseed meal and explore its cotton meal detoxification effect.【Results】 It was found that the optimal temperature of strain JD-1 was 37℃, and with the extension of time, the growth concentration of the strain gradually increased and stabilized in 22-24 h. However, the pH of the JD-1 strain gradually decreased with time, and finally stabilized. With the increase of the salt concentration within 24 h, the growth of the strain gradually slowed down until it finally stabilized, and it had a certain salt tolerance. But too much acid or too much alkali would inhibit its growth. Under optimal conditions, the degradation rate of JD-1 fermented cottonseed meal to reduce gossypol was 40%, and its evolutionary relationship was as high as 99% homology with Pediococcus lactis. It was determined that JD-1 strain was lactic acid Pediococcus.【Conclusion】 In this paper, a strain of Lactococcus lactis with good ability to degrade gossypol was reported with 40% detoxification of cotton phenol from cotton meal, using gossypol as the sole carbon source.

Key words: free gossypol; strain screening; gossypol detoxification; biological characteristics

中图分类号:

S188

张莉, 杨晓龙, 王海亮, 南珊珊, 马雪儿, 聂存喜, 张文举, 陈程. 一株棉酚降解菌株的筛选鉴定及对棉粕的脱毒效果[J]. 新疆农业科学, 2022, 59(12): 3057-3065.

ZHANG Li, YANG Xiaolong, WANG Hailiang, NAN Shanshan, MA Xueer, NIE Cunxi, ZHANG Wenju, CHEN Cheng. Selection and Identification of a Cotton Phenol Degradation Strain and Its Detoxification Effect on Cotton Meal[J]. Xinjiang Agricultural Sciences, 2022, 59(12): 3057-3065.

图/表 9

表1 试验组条件

Table 1 Test group conditions

项目 Project	温度 Temperature (℃)	时间 Time (h)	添加量 Addition (%)	底物水份 Substrate moisture (%)
乳酸片球菌组Pediococcus lactis	37	48	5	40
热带假丝酵母ZD-3组Candida tropicalis ZD-3 group	30	48	5	40

图1 24 h内JD-1在不同温度的生长状态

Fig.1 The growth state of JD-1 at different temperatures within 24 h

图2 24 h内最适温度下JD-1生长曲线的绘制

Fig.2 The growth curve of JD-1 at the optimum temperature within 24 h

图3 24 h内最适温度下JD-1 pH的变化

Fig.3 The change of JD-1 PH value at the optimum temperature within 24 h

图4 24 h内最适温度下JD-1的耐盐情况

Fig.4 Salt tolerance of JD-1 at the optimum temperature within 24 h

图5 24 h内最适温度下JD-1的酸碱耐受情况

Fig.5 The acid-base tolerance of JD-1 at the optimum temperature within 24 hours

表2 不同处理对棉籽粕中FG的含量和脱毒率

Table 2 Different treatments on the FG content in cottonseed meal

分组 Group	FG含量 FG content (mg/kg)	脱毒率 Detoxification rate (%)
对照组(只高压未发酵) Control group (only high- pressure unfermented)	98.06±1.38^b	79
棉粕原料组(未处理) Cotton meal raw material group (untreated)	485.15±8.93^a	*
乳酸片球菌组 Pediococcus lactis	59.45±1.11^c	40
热带假丝酵母ZD-3组 Candida tropicalis ZD-3 group	50.47±1.30^c	48

图6 JD-1革兰氏染色

Fig.6 JD-1 Gram staining microscope image

图7 JD-1系统发育树

Fig.7 JD-1 phylogenetic tree

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一株棉酚降解菌株的筛选鉴定及对棉粕的脱毒效果

Selection and Identification of a Cotton Phenol Degradation Strain and Its Detoxification Effect on Cotton Meal

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