天津农学院学报 ›› 2018, Vol. 25 ›› Issue (3): 60-64.doi: 10.19640/j.cnki.jtau.2018.03.013

• 研究与简报 • 上一篇    下一篇

L-色氨酸重组高产菌株发酵工艺的优化

吕磊1, 宋国田2, 刘永飞2, 刘燕霏1, 杨建德1,通信作者, 张大伟2,通信作者   

  1. 1. 天津农学院 动物科学与动物医学学院,天津300384;
    2. 中国科学院 天津工业生物技术研究所,天津300380
  • 收稿日期:2018-03-29 出版日期:2018-09-20 发布日期:2019-11-12
  • 通讯作者: 杨建德(1969-)男,教授,博士,研究方向:预防兽医学。E-mail:jiandeyang@126.com。张大伟(1978-),男,研究员,博士,研究方向:微生物发酵。E-mail:zhang _dw@tib.cas.cn。
  • 作者简介:吕磊(1995-),男,本科在读,研究方向:微生物发酵。E-mail:824951764qq.com。
  • 基金资助:
    天津自然科学基金项目(16JCYBJC23500)

Optimization of fermentation process for recombinant high yield strain of L-tryptophan

LÜ Lei1, SONG Guo-tian2, LIU Yong-fei2, LIU Yan-fei1, YANG Jian-de1,Corresponding Author, ZHANG Da-wei2,Corresponding Author   

  1. 1. College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300384, China;
    2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Science, Tianjin 300380, China
  • Received:2018-03-29 Online:2018-09-20 Published:2019-11-12

摘要: L-色氨酸(L-tryptophan,L-Trp)是一种限制性氨基酸,是人体和动物体生长发育等重要生理活动的必需氨基酸之一,也是合成许多活性物质的关键氨基酸之一,广泛应用于医药、饲料和食品等方面,具有十分重要的商业价值。微生物发酵生产的L-色氨酸近些年来一直备受关注,主要是因为其价格低廉、纯度高、易分离等特点,因此利用微生物生产L-色氨酸在大规模工业生产上具有十分可观的应用前景。利用色氨酸高产基因,经基因工程重组到大肠杆菌内,筛选出色氨酸高产菌株。通过控制溶氧、碳源、pH值及残糖含量来有效提高L-色氨酸的产量。利用原始菌株探究发酵过程中发酵罐内的残糖含量,发现残糖含量控制在1% 时,菌体生长最为良好,生长周期最长。将不同比例的糖蜜与葡萄糖混合,作为发酵时的碳源,结果显示当比例为3:1时L-色氨酸产量最大;在发酵过程中控制pH条件,将产量提高到18.9 g/L;将溶氧条件控制在35%、40% 两个范围,结果显示溶氧40% 时L-色氨酸的产量达到27 g/L;将补料碳源换为60% 的葡萄糖及70% 的小麦糖,结果显示葡萄糖作为唯一碳源时,L-色氨酸产量较高。

关键词: L-色氨酸, 发酵, 优化, 高产菌株

Abstract: L-tryptophanine(L-Trp)is a kind of restricted amino acid, which is one of the essential amino acids for the growth and development of human body and animal. It is one of the key amino acids in the synthesis of many active substances and has been widely used in medicine. The production of L- tryptophan by microbial fermentation has attracted much attention in recent years, mainly because of its low price, high purity and easy separation. Therefore, the production of L-tryptophan by microorganism has a promising application prospect in large-scale industrial production. In this paper, the tryptophan high yield gene was recombined into Escherichia coli by genetic engineering. By controlling dissolved oxygen, pH value of carbon source and residual sugar content, the yield of L-tryptophan was effectively increased by direct fermentation. In this experiment, the original strain was used to explore the content of residual sugar in fermenting tank. It was found that when the residual sugar content was controlled at 1%, the growth of bacteria was the best and the growth cycle was the longest. When molasses were mixed with glucose as carbon source during fermentation, the results showed that the yield of L-tryptophan was the highest when the ratio was 3:1; then when the pH was controlled during fermentation, the yield was increased to 18.9 g/L;the dissolved oxygen condition was controlled in the range of 35% and 40%. The results showed that the production of L-tryptophan reached 27 g/L at dissolved oxygen 40%; Finally, 60% glucose and 70% wheat sugar were replaced by carbon source. The results showed that the yield of L-tryptophan was higher when glucose was the sole carbon source.

Key words: L-tryptophan, fermentation, optimization, high yield strain

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