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        Development of a High-Titer Culture Medium for the Production of Cholesterol by Engineered Saccharomyces cerevisiae and Its Fed-Batch Cultivation Strategy

        Wang, Ling-Xu,Zheng, Gao-Fan,Xin, Xiu-Juan,An, Fa-Liang The Korean Society for Microbiology and Biotechnol 2022 Journal of microbiology and biotechnology Vol.32 No.9

        Steroids are a class of compounds with cyclopentane polyhydrophenanthrene as the parent nucleus, and they usually have unique biological and pharmacological activities. Most of the biosynthesis of steroids is completed by a series of enzymatic reactions starting from cholesterol. Synthetic biology can be used to synthesize cholesterol in engineered microorganisms, but the production of cholesterol is too low to further produce other high-value steroids from cholesterol as the raw material and precursor. In this work, combinational strategies were established to increase the production of cholesterol in engineered Saccharomyces cerevisiae RH6829. The basic medium for high cholesterol production was selected by screening 8 kinds of culture media. Single-factor optimization of the carbon and nitrogen sources of the culture medium, and the addition of calcium ions, zinc ions and citric acid, further increased the cholesterol production to 192.53 mg/l. In the 5-L bioreactor, through the establishment of strategies for glucose and citric acid feeding and dissolved oxygen regulation, the cholesterol production was further increased to 339.87 mg/l, which was 734% higher than that in the original medium. This is the highest titer of cholesterol produced by microorganisms currently reported. The fermentation program has also been conducted in a 50-L bioreactor to prove its stability and feasibility.

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        De novo assembly and transcriptome analysis of differentially expressed genes relevant to variegation in hawthorn flowers

        Wei Ji,Wei Zhao,Rong‑Chen Liu,Xiao‑Bo Jiao,Kai Han,Zhong‑Yi Yang,Mei‑Ying Gao,Rui Ren,Xiu‑Juan Fan,Ming‑Xia Yang 한국식물생명공학회 2019 Plant biotechnology reports Vol.13 No.6

        Flower color variegation has been observed in many plant species. However, pink flowers on the white-blooming hawthorn trees found by our group earlier have never been reported. To better understand the differentially expressed genes (DEGs) in variegated hawthorn flowers, white and pink flowers at different developmental stages (S1 and S2) underwent transcriptome sequencing separately. Approximately 34.28 Gb of high-quality data were obtained and assembled into 100,013 unigenes with an average length of 706.93 bp. These unigenes were further subjected to functional annotation and biochemical pathway analysis, and DEGs of two types of hawthorn flowers at different developmental stages were studied. Based on the enrichment analysis of DEGs, eight anthocyanin-modified enzyme genes or other enzyme genes that indirectly affect anthocyanin synthesis (5AT, 3GGT , and AI, β-Glu, two Aux/IAAs, two PODs), eight structural genes (UFGT, DFR, CHI, two F3Hs, and three PALs), and three transcription factors (one MYB and two bHLHs) were also identified. We randomly selected 15 genes, and the trends in the expression levels of these genes in the organs of white and pink flowers at different developmental stages were verified by quantitative real-time PCR. Mass sequence data obtained by RNA-seq of variegated hawthorn flowers provided basic sequence information and a unique opportunity to uncover the genetic mechanisms under-lying flower color variegation.

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