秸秆糖发酵制备琥珀酸及菌种选育

李兴江; 姜绍通; 潘丽军

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秸秆糖发酵制备琥珀酸及菌种选育

首发时间：2009-01-29

李兴江 ¹ 姜绍通 ¹ 潘丽军 ¹

1、合肥工业大学生物与食品工程学院

摘要：在产琥珀酸放线杆菌主要酶活分析基础上选育出高产突变株对工业规模生物转化农作物秸秆制备琥珀酸具有重要意义。采用酸解方法处理原料秸秆并利用高效液相检测降解糖分，在菌株主要酶活分析及调控的基础上，利用软X射线诱变结合丙烯醇抗性平板进行定向选育以降低乙醇流量，并对代谢酶活进行了分析。秸秆水解液的检测表明200g的秸秆干粉酶解得约47g的葡萄糖和约26g的木糖，酶活分析表明Pepck、Pc及Mdh是琥珀酸合成重要代谢酶且酶活可调控，发现Adh的高酶活造成了大量副产物乙醇的生成。筛选出的丙烯醇抗性突变株S.JST01的乙醇产量由8.9g/L降为1.4g/L，降低了84%，而琥珀酸流量产量由54.2g/L升至63.1g/L，升高了16%。酶活检测表明乙醇脱氢酶（Adh）的酶活力单位从614降低到108。围绕产琥珀酸放线杆菌Adh酶活降低实施定向选育有助于提高琥珀酸的流量，突变株的末端产物琥珀酸累积达63.1g/L，具有工业应用潜力。

关键词：农作物秸秆产琥珀酸放线杆菌乙醇脱氢酶丙烯醇筛选

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The fermentation of Actinobacillus succinogenes with crop straw hydrolysate and its screening

lixingjiang ¹ jiangshaotong ² panlijun ²

1、School of Biotechnology and Food Engineering, Hefei University of Technology
2、School of Biotechnology and Food Engineering, Hefei University of Technology,

Abstract：It is very important to obtain high yield mutant strain on the base of key enzyme activity analysis of A.succinogenes for the industrial bioconversion of succinic acid from crop straw. The pentose and hexose hydrolyzed from crop straw with dilute sulfuric acid were determinated by HPLC. During the fermentation of the hydrolysate the activity of key enzymes were determinated and regulated. In order to decrease ethanol yield,those strains mutated by soft X-ray of synchronous radiation were screened out on the plates with high concentration of allyl alcohol. Then the alcohol dehydrogenase activity of the mutant strain was compared with that of the parent strain. Determination of the hydrolysate showed that obout 47g glucose and 26g xylose were hydrolyzed from 200g crop straw. Pepck and Pc and Mdh were identified as the key enzymes of succinic acid metabolism and the high activity of Adh caused the cumulation of ethanol. Determination of end products from the parent strain indicated that the concentration of ethanol was highest among those byproducts and that the yield of product succinic acid and byproduct ethanol were 54.2g/L and 8.9g/L ,respectively. Compared with the parent strain,the ethanol concentration of anti-allyl-alcohol mutant strain S.JST01 decreased by 84%，from 8.9g/L to 1.4g/L. Meanwhile the yield of succinic acid increased by 16%, from 54.2g/L to 63.1g/L. The enzyme determination showed that the activity unit of alcohol dehydrogenase decreased from 614 to 108. The interdiction of metabolic pathway of Adh decreased the metabolism of ethanol accordingly, thus the succinic acid flux was strengthened by the redundant carbon flux from the byproduct. And the mutant strain S.JST01 with the end product yield of 63.1g/L succinic acid deserves being extended to application of industrial fermentation.

Keywords： crop straw, Actinobacillus succinogenes, alcohol dehydrogenase, allyl alcohol, screening

基金：

1. 教育部博士点基金,国家科技支撑计划,安徽省自然科学基金,合肥工业大学科学研究发展基金（200803590001,2007BAD34B01,070411024,073002F）

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李兴江，姜绍通，潘丽军. 秸秆糖发酵制备琥珀酸及菌种选育[EB/OL]. 北京：中国科技论文在线 [2009-01-29]. https://www.paper.edu.cn/releasepaper/content/200901-1160.

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论文编号	200901-1160
论文题目	秸秆糖发酵制备琥珀酸及菌种选育
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