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中国生物工程杂志

China Biotechnology
中国生物工程杂志  2017, Vol. 37 Issue (12): 40-48    DOI: 10.13523/j.cb.20171207
研究报告     
HOG1抑制剂调节球头三型孢菌多元醇生产及机理 *
顾丽娜1,李良智1,2(),郭伟强1,顾竟生1,姚雪梅1,鞠鑫1
1 苏州科技大学化学生物与材料工程学院 苏州 215009
2 新加坡国立大学生物科学系 新加坡 117543
The Regulation on Polyols Production by Trichosporonoides oedocephalis with HOG1 Inhibitors and Its Mechanism
Li-na GU1,Liang-zhi LI1,2(),Wei-qiang GUO1,Jing-sheng GU1,Xue-mei YAO1,Xin JU1
1 School of Chemistry, Biology, and Materials Engineering, Suzhou University of Science and Technology,Suzhou 215009, China
2 Department of Biological Sciences, National University of Singapore, Singapore 117543
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摘要: 目的

采用HOG1抑制剂对球头三型孢菌产多元醇进行调控。

方法

向培养基中加入SB239063、SB202190和SB203580三种抑制剂进行发酵实验,比较三种抑制剂对发酵的影响。

结果

实验结果表明SB239063可以降低球头三型孢菌细胞内胞浆3-磷酸甘油脱氢酶(ctGPD)的酶活,提高赤藓糖还原酶(ER)的酶活。此外对HOG1和Phospho-HOG1的Western blot结果分析显示,SB239063还会抑制球头三型孢菌细胞内HOG1的脱磷酸化。最终添加10μmol/L SB239063使发酵120 h后的多元醇产物中甘油产量下降20.57%,赤藓糖醇产量提高31.16%,底物转化率提高24.73%。

结论

SB239063可以降低球头三型孢菌产甘油的能力,提高赤藓糖醇的产量。

关键词: 球头三型孢菌HOG1抑制剂赤藓糖醇赤藓糖还原酶    
Abstract: Objective:

To regulate the production of polyols by Trichosporonoides oedocephalis employing HOG1 inhibitors.

Methods:

SB239063, SB202190 and SB203580 were added into the medium to conduct fermentation experiments, and the influence of three inhibitors on the fermentation was compared.

Results:

The experimental results showed SB239063 would decrease the ctGPD enzyme activity of Trichosporonoides oedocephalis, and enhance erythritol reductase(ER)enzyme activity at the same time.Furthermore, the Western blot analysis of HOG1 and phospho-HOG1demonstrated that SB239063 also inhibited the dephosphorylation of HOG1. After 120 h fermentation, the addition of 10μmol/L SB239063 decreased glycerol production by 20.57% and increased erythritol production by 31.16%.In other words, the conversion rate of glucose was increased by 24.73%.

Conclusion:

SB239063 weakened the ability to produce glycerol of Trichosporonoides oedocephalis and therefore promoted erythritol yield.

Key words: Trichosporonoides oedocephalis    HOG1 inhibitors    Erythritol    Erythritol reductase
收稿日期: 2017-05-29 出版日期: 2017-12-16
ZTFLH:  Q789  
基金资助: 国家自然科学基金(21376156)、江苏省高校“青蓝工程”资助项目
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顾丽娜
李良智
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姚雪梅
鞠鑫

引用本文:

顾丽娜,李良智,郭伟强,顾竟生,姚雪梅,鞠鑫. HOG1抑制剂调节球头三型孢菌多元醇生产及机理 *[J]. 中国生物工程杂志, 2017, 37(12): 40-48.

Li-na GU,Liang-zhi LI,Wei-qiang GUO,Jing-sheng GU,Xue-mei YAO,Xin JU. The Regulation on Polyols Production by Trichosporonoides oedocephalis with HOG1 Inhibitors and Its Mechanism. China Biotechnology, 2017, 37(12): 40-48.

链接本文:

http://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20171207        http://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I12/40

图1  HOG1抑制剂对球头三型孢菌发酵的影响
图2  添加HOG1抑制剂的发酵时间曲线
图3  添加HOG1抑制剂形成的抑菌圈
Time (h) control SB202190 SB203580 SB239063
24 100 99 ± 2 96 ± 3 90 ± 1
48 100 92 ± 1 94 ±2 96 ± 3
72 100 96 ± 2 87 ± 1 80 ± 1
96 100 95 ± 1 86 ± 1 81 ± 1
120 100 94 ± 1 92 ± 2 87 ± 2
表1  MTT法测定球头三型孢菌细胞活性(%)
图4  HOG1和Phospho-HOG1的Western blot 分析
发酵时间(h) Specificactivity of ctGPD(mU/mg protein)
control SB202190 SB203580 SB239063
24 18.67 ± 0.29 18.37 ± 0.32 11.55 ± 0.25 15.03 ± 0.31
48 17.58 ± 0.26 17.33 ± 0.17 7.79 ± 0.09 14.23 ± 0.28
72 6.99 ± 0.18 6.64 ± 0.36 5.85 ± 0.12 6.12 ± 0.13
96 4.28 ± 0.16 4.18 ± 0.24 5.43 ± 0.11 4.05 ± 0.08
120 3.26 ± 0.13 3.23 ± 0.19 3.47 ± 0.08 3.17 ± 0.07
表2  HOG1抑制剂对ctGPD酶活的影响
发酵时间(h) Specificactivity of ER(mU/mg protein)
control SB202190 SB203580 SB239063
24 327.59 ± 10.23 240.33 ± 7.39 251.57 ± 10.31 278.03 ± 11.21
48 227.63 ± 8.76 180.77 ± 6.29 167.83 ± 2.78 196.58 ± 9.73
72 126.52 ± 7.82 127.79 ± 6.34 129.26 ± 2.81 168.53 ± 5.27
96 104.49 ± 5.43 108.12 ± 5.76 110.41 ± 1.98 136.32 ± 4.19
120 87.34 ± 3.86 78.93 ± 3.21 80.13 ± 1.76 92.73 ± 2.17
表3  HOG1抑制剂对ER酶活的影响
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