《中药生物技术》教学：BTTCM8

赵秀举

摘要

一，复习和检查预习

二，冬虫夏草菌丝体的液体深层发酵培养

三，课外拓展（自由选择，有时候选择比努力重要！）

《中药生物技术》教学进度

教材： 中药生物技术，贾景明，专业教材，化学工业出版社，2008

一，复习和检查预习

1，灵芝属于            科。

2，[微生物]发酵的分类标准有：                、               、                       、                       。

3，表4-3真菌制剂基源的活性成分。一到五组，分别是1、2，3、4，7、8，9、5，11、6.

二，授课

发酵技术的过程

冬虫夏草菌丝体的液体深层发酵培养

三，课外拓展/翻译提示

Investigation of liquid–solid hydrodynamic boundary layers and oxygen requirements in hairy root cultures. Gary R. C. Williams, Pauline M. Doran* Biotechnol Bioeng, 1999, 64:729-740

Biotechnology and Bioengineering

(Biotechnology & Applied Microbiology)

Keywords（关键词）:

Atropa belladonna（颠茄）; critical oxygen tension（临界氧分压）; hair roots（毛状根/发根）;

mucilage（黏液）; oxygen mass transfer（氧传递）; Solanum aviculare（澳洲茄）[Solanum melongena茄子]

Abstract（摘要）

Rates of oxygen uptake, growth and alkaloid production by hairy roots in submerged culture were investigated using a recirculation reactor allowing operation at high liquid velocities for removal of hydrodynamic boundary layers.

毛状根深层培养时的摄氧率、生长率和生物碱产生率利用再循环生物反应器进行调查，这种反应器在高液速时移除流动分界层。[图1]

Measurements were performed at dissolved oxygen tensions of 31–450% air saturation.

测量在溶解氧分压为31 -450%空气饱和度的条件下进行。

Critical oxygen concentrations for Atropa belladonna hairy roots were above airsaturation, viz.   [videlicet] 100–125% air saturation for oxygen uptake and 150% air saturation for growth, demonstrating that these roots cultivated in reactors with air sparging are oxygen-limited.

颠茄毛状根培养的临界氧浓度高于空气饱和度,即摄氧时100 - 125%空气饱和度，生长时150%空气饱和度,表明这些在空气喷射反应器中进行的毛状根培养，氧气为限制因素。[图3]

The critical oxygen tension for oxygen uptake by Solanum aviculare hairy roots was 75% air saturation.

澳洲茄毛状根摄氧时的临界氧分压是75%空气饱和度。[图5]

 Both the specific oxygen uptake rate and specific growth rate of A. belladonna hairy roots were dependent on the mass (g dry weight) of roots present; even in the absence of boundary layers, growth did not remain exponential over the entire culture period.

颠茄毛状根的比摄氧率和比生长速率均取决于当时的质量(克干重);即使没有边界层,生长也不会在整个培养时期保持指数状态。[图4、7]

Cryo-scanning electron microscopy showed that hairy roots grown submerged in liquid mediumwere covered with thick layers of hydrated mucilage and root hairs, representing a significant additional barrier to oxygen transfer.

低温扫描电子显微镜显示,液体培养基深层培养时，生长的毛状根覆盖着水化黏液厚层和根发是 ,氧转移过程一个重要的附加障碍。[图9]

Roots protruding out of the liquid medium showed no evidence of mucilage accumulation.

而生长在液体培养基外的毛状根则没有黏液积聚。

The specific oxygen demand of A. belladonna root tips was 3.3–11.5 times higher than for the remainder of the roots, the ratio increasing as the dissolvedoxygen tension was reduced.

颠茄根尖比需氧量是根其余部位的3.3 - -11.5倍,且随着溶氧分压降低这一比率上升。[图6]

Specific growth rates, biomass yields from sugar, and atropine levels were maximum at around 150% air saturation, but decreased significantly with oxygen concentrations above ca. 200%.

比生长率、糖和阿托品生物量在150%空气饱和度下达到最高,但当氧气浓度在大约200%空气饱和度以上时显著降低。[图8]

四，作业

计算（10分）中药活性成分代谢/发酵技术

请根据表1和2，确定固态发酵生产药物MK的最优组合。

表1 固态发酵条件优化正交试验因素与水平

水平  A 高温时长/d  B 低温时长/d  C装料量/g  D （空，误差项）

1  3  12  300

2  5  15  360

3  6  17  500

表2 固态发酵条件优化正交试验结果

试验 A B C D MK/‰

1 1 1 1 1 10

2 1 2 2 2 17

3 1 3 3 3 11

4 2 1 3 2 13

5 2 2 1 3 17

6 2 3 2 1 20

7 3 1 2 3 18

8 3 2 3 1 13

9 3 3 1 2 15

[选自“赵秀举*, 刘志国. 红曲洛伐他汀发酵条件优化及降脂功能. 中国酿造，2014, 33(1), 32-35. ”]

往期回顾

《中药生物技术》课程教学体会  http://blog.sciencenet.cn/blog-392245-640411.html

"课程与就业"微信公众号