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Carbohydrate analysis of Mortierella alpina by colorimetry and HPLC–ELSD to reveal accumulation differences of sugar and lipid
Biotechnology Letters ( IF 2.7 ) Pub Date : 2021-04-17 , DOI: 10.1007/s10529-021-03120-2
Hanqin Chen 1, 2 , Haiqin Chen 1, 2 , Hengqian Lu 1, 2 , Xin Tang 1, 2 , Hao Zhang 1, 2, 3, 4 , Yong Q Chen 1, 2, 3, 4 , Wei Chen 1, 2, 3, 5
Affiliation  

Objectives

To establish reliable methods for the extraction and quantification of the total carbohydrate and intracellular saccharides from Mortierella alpina and study the changes between carbohydrate and lipid in fermentation process.

Results

The extraction of mycelia with HCl following a photometric phenol–sulphuric acid reaction was identified as an optimal method for total carbohydrate analysis in Mortierella alpina, which the extraction efficiency performed 1.1–3.6 fold than other five methods. The total carbohydrate content increased from initial 19.26 to 25.86% during early fermentation process and declined gradually thereafter, while the fatty acid was increasing from 8.47 to 31.03%. For separation and qualitative estimation of intracellular saccharides, the acetonitrile/water freeze–thaw method for extraction and Sugar-Pak I column for separation proved to be possible. With the glucose rapidly decreasing at the beginning of growth, the trehalose accumulated rapidly from 1.63 to 5.04% and then decreased slightly but maintain above 4% of dry biomass.

Conclusions

This work established comprehensive carbohydrate extraction and analysis methods of Mortierella alpina and identified the main saccharide in fermentation process which indicated that the accumulation of fatty acids was related to the change of intracellular carbohydrate content.



中文翻译:

通过比色法和 HPLC-ELSD 分析高山被孢霉的碳水化合物以揭示糖和脂质的积累差异

目标

建立可靠的高山被孢霉总糖类和胞内糖类的提取和定量方法,研究发酵过程中糖类和脂质的变化。

结果

在光度法苯酚-硫酸反应后用 HCl 提取菌丝体被确定为高山被孢霉总碳水化合物分析的最佳方法,其提取效率是其他五种方法的 1.1-3.6 倍。总碳水化合物含量在早期发酵过程中从最初的19.26%增加到25.86%,之后逐渐下降,而脂肪酸从8.47%增加到31.03%。对于细胞内糖类的分离和定性评估,乙腈/水冻融提取法和 Sugar-Pak I 柱分离被证明是可行的。随着生长初期葡萄糖的迅速下降,海藻糖从1.63%迅速积累到5.04%,然后略有下降,但保持在4%以上的干生物量。

结论

本工作建立了高山被孢霉碳水化合物的综合提取和分析方法,并确定了发酵过程中的主要糖类,表明脂肪酸的积累与细胞内碳水化合物含量的变化有关。

更新日期:2021-04-18
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