嗜热链球菌AR333胞外多糖合成的工程udp -糖合成途径

IF 4 2区农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY Journal of the Science of Food and Agriculture Pub Date : 2025-03-03 DOI:10.1002/jsfa.14206

Yizhou Fan, Junkang Shi, Xin Song, Guangqiang Wang, Yongjun Xia, Lianzhong Ai, Zhiqiang Xiong

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引用次数: 0

摘要

背景：嗜热链球菌（Streptococcus thermoophilus）产生的胞外多糖（Exopolysaccharide， EPS）能显著改善乳制品的粘度、质地和口感，在食品工业中具有广阔的应用潜力。然而，嗜热链球菌的EPS产量通常很低（-1），难以满足工业化要求。利用代谢工程技术对嗜热链球菌进行基因改造是促进EPS生物合成的有效途径。结果：据我们所知，目前还缺乏对嗜热葡萄球菌生产EPS的工程udp -糖合成途径的系统研究。在本研究中，通过调控EPS前体udp -葡萄糖和udp -半乳糖的生物合成基因来提高嗜热链球菌AR333的EPS产量。与对照相比，单独过表达8个EPS前体基因的工程菌株EPS产量分别提高了7-31%。其中，编码葡萄糖激酶的glk和编码udp -半乳糖-4-epimase的galE1过表达，EPS产量分别为275.37 mg L-1和288.65 mg L-1。此外，编码β-半乳糖苷酶和galE1的lacZ共过表达使EPS产量显著增加49% （329.51 mg L-1）。转录分析进一步表明，工程菌株EPS合成的增强可能与EPS基因的前体基因和簇的上调有关。结论：我们的研究结果表明，工程化的udp -糖合成是提高嗜热葡萄球菌EPS产量的有效策略。©2025化学工业协会。

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Engineering UDP-sugar synthesis pathways for exopolysaccharide biosynthesis in Streptococcus thermophilus AR333

BACKGROUND

Exopolysaccharide (EPS) produced by Streptococcus thermophilus can significantly improve the viscosity, texture and taste of dairy products, which have broad application potential in the food industry. However, EPS production is typically low (< 100 mg L⁻¹) in S. thermophilus, making it difficult to meet industrialization requirements. The utilization of metabolic engineering for genetic modification of S. thermophilus is an efficient approach to enhance EPS biosynthesis.

RESULTS

To our knowledge, there is lack of systematic investigation on engineering UDP-sugar synthetic pathways for EPS production in S. thermophilus. In the present study, the biosynthetic genes of EPS precursors UDP-glucose and UDP-galactose were regulated for improving EPS production in S. thermophilus AR333. Compared with the control, engineered strains by single overexpression of eight EPS precursor genes increased EPS production by 7–31%, respectively. Among of them, overexpressing glk encoding glucokinase and galE1 encoding UDP-galactose-4-epimase led to 275.37 and 288.65 mg L⁻¹ of EPS production, respectively. Moreover, co-overexpression of lacZ encoding β-galactosidase and galE1 achieved a remarkable 49% increase in the EPS production (329.51 mg L⁻¹). Transcriptional analysis further suggested that enhanced EPS synthesis in engineered strain can attributed to the upregulation of precursor genes and clusters of EPS genes.

CONCLUSION

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来源期刊

Journal of the Science of Food and Agriculture 工程技术-农业综合

CiteScore

8.10

自引率

4.90%

发文量

634

审稿时长

3.1 months

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