Safety evaluation and optimization of high-efficiency expression of 5hsCT in Saccharomyces cerevisiae.

IF 2.1 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2025-06-14 DOI:10.1007/s10529-025-03607-2

Yakun Zhang, Wenlu Jia, Wenli Qiu, Yujing Tang, Hualin Wei, Hengyi Sun

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Abstract

To evaluate the oral safety of the industrial strain Saccharomyces cerevisiae YS-5hsCT (S. cerevisiae YS-5hsCT) expressing human/salmon chimeric calcitonin (5hsCT) and optimize its fermentation conditions, providing scientific basis for industrial production of oral recombinant calcitonin. The strain demonstrated an LD₅₀ > 10 g kg⁻¹, with all toxicological tests yielding negative results and normal hematological/biochemical parameters (P > 0.05). mRNA analysis revealed optimal gene expression at 96 h, 30 °C and 225 rpm. Single-factor experiments established optimal growth conditions: 8% initial inoculum size, 72 h culture, 30 °C, 200-250 rpm agitation, and 7-8.5 l min⁻¹ aeration. Under optimized conditions (10% inoculum size, 72 h, 25 °C, 225 rpm, 7 l min⁻¹ aeration), the biomass reached 7.175 g l⁻¹ d⁻¹ and the 5hsCT protein yield was 149.24 mg l⁻¹ d⁻¹. These findings confirm the oral safety of YS-5hsCT and providetechnical support for industrial calcitonin production. Safety evaluations demonstrated transgenic S. cerevisiae YS-5hsCT exhibits no toxicity or mutagenicity, with optimized cultivation conditions enhancing biomass yield for potential calcitonin production applications.

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5hsCT在酿酒酵母中高效表达的安全性评价及优化。

目的评价表达人/鲑鱼嵌合降钙素（5hsCT）的工业菌株酿酒酵母YS-5hsCT （S. cerevisiae YS-5hsCT）的口服安全性，并优化其发酵条件，为口服重组降钙素的工业化生产提供科学依据。毒株的毒理学测试结果为阴性，血液学/生化参数正常（P > 0.05）。mRNA分析显示，在96 h， 30°C和225 rpm时基因表达最佳。单因素实验确定了最佳生长条件：8%的初始接种量，72小时培养，30°C， 200-250转搅拌，7-8.5分钟曝气。在最佳条件下（10%接种量，72 h, 25°C， 225转/分，7 l分钟毒血症），生物量达到7.175 g l -毒血症，5hsCT蛋白产量为149.24 mg l -毒血症。这些结果证实了YS-5hsCT的口服安全性，为降钙素的工业化生产提供了技术支持。安全性评估表明，转基因酿酒葡萄球菌YS-5hsCT没有毒性或突变性，优化的培养条件提高了生物产量，可用于降钙素的生产。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.