Accumulation of squalene in a microalga Chlamydomonas reinhardtii by genetic modification of squalene synthase and squalene epoxidase genes

PloS One
Masataka KajikawaHideya Fukuzawa

Abstract

Several microalgae accumulate high levels of squalene, and as such provide a potentially valuable source of this useful compound. However, the molecular mechanism of squalene biosynthesis in microalgae is still largely unknown. We obtained the sequences of two enzymes involved in squalene synthesis and metabolism, squalene synthase (CrSQS) and squalene epoxidase (CrSQE), from the model green alga Chlamydomonas reinhardtii. CrSQS was functionally characterized by expression in Escherichia coli and CrSQE by complementation of a budding yeast erg1 mutant. Transient expression of CrSQS and CrSQE fused with fluorescent proteins in onion epidermal tissue suggested that both proteins were co-localized in the endoplasmic reticulum. CrSQS-overexpression increased the rate of conversion of 14C-labeled farnesylpyrophosphate into squalene but did not lead to over-accumulation of squalene. Addition of terbinafine caused the accumulation of squalene and suppression of cell survival. On the other hand, in CrSQE-knockdown lines, the expression level of CrSQE was reduced by 59-76% of that in wild-type cells, and significant levels of squalene (0.9-1.1 μg mg-1 cell dry weight) accumulated without any growth inhibition. In co-transformation lines...Continue Reading

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Citations

Dec 24, 2015·Frontiers in Microbiology·Javier A GimpelStephen P Mayfield
Mar 11, 2017·Biotechnology Advances·Xiangzhao MaoSang Yup Lee
Apr 21, 2018·Critical Reviews in Food Science and Nutrition·Ming-Hua LiangJian-Guo Jiang
Oct 13, 2016·World Journal of Microbiology & Biotechnology·Wen XuYang Wang
Apr 11, 2019·Frontiers in Bioengineering and Biotechnology·Nisarg GohilVijai Singh
Jun 28, 2019·Plant Direct·Kristina UnlandChristian Schulze Gronover
Oct 6, 2020·Frontiers in Bioengineering and Biotechnology·Gulshan KumarTilak Raj Sharma
Jan 22, 2021·FEMS Microbiology Ecology·Isiri Adhiwarie PereraMallavarapu Megharaj
Jun 15, 2021·Applied Microbiology and Biotechnology·Peng-Wei HuangHe Huang
Aug 29, 2021·Plants·Supakorn PotijunAnchalee Sirikhachornkit

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Methods Mentioned

BETA
genetic modification
electrophoresis
transgenic
PCR

Software Mentioned

SOSUI
MicorRNA Designer

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