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阻断消耗途径提高毕赤酵母工程菌S-腺苷甲硫氨酸产量
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国家自然科学基金(31300076);广西壮族自治区自然科学基金(2019GXNSFAA245001,2018GXNSFAA281005,2017GXNSFAA198136)


Blocking consumption pathway increases production of S-adenosyl-l-methionine by Pichia pastoris
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    摘要:

    【背景】S-腺苷甲硫氨酸(S-adenosyl-l-methionine, SAM)作为所有生物体内的重要中间代谢物,不仅可作为膳食补充剂,还具有良好的临床应用价值。【目的】将毕赤酵母重组菌GS115/DS16的SAM消耗途径阻断,进一步提高SAM的产量。【方法】分别敲除毕赤酵母重组菌GS115/DS16的S-腺苷同型半胱氨酸水解酶基因sah1、S-腺苷甲硫氨酸脱羧酶基因spe2和l-甲硫氨酰tRNA合酶基因msm1,构建工程菌G/Dsah、G/Dspe和G/Dmsm。检测3个工程菌的生长和SAM产量,以及l-Met添加量对SAM积累的影响。【结果】与出发菌GS115/DS16相比,工程菌G/Dsah、G/Dspe和G/Dmsm的单位菌体SAM产量分别提高了29.3%、55.6%和24.8%,其生长无显著差异。l-Met添加量优化后(0.06%),G/Dsah和G/Dmsm单位菌体的SAM产量分别提高了26.4%和28.9%。【结论】构建的毕赤酵母工程菌可用于SAM的工业化生产,该代谢工程策略可用于改进其他化学品的生产。

    Abstract:

    [Background] S-Adenosyl-l-methionine (SAM) is an important intracellular metabolite that can be used as a dietary supplement and to treat a variety of diseases. [Objective] To enhance SAM production for industrial application by blocking SAM consumption pathways in the recombinant Pichia pastoris strain GS115/DS16. [Methods] The genes associated with the metabolism of SAM, sah1 (encoding S-adenosyl-l-homocysteine hydrolase), spe2 (encoding S-adenosylmethionine decarboxylase), and msm1 (encoding mitochondrial methylthio-tRNA synthase), were knocked out in SAM-producing strain GS115/DS16. Accordingly, the engineered strains G/Dsah, G/Dspe, and G/Dmsm were constructed. The cell growth and SAM production of the three engineered strains were investigated. Additionally, the effect of methionine addition on SAM accumulation was studied. [Results] The knock-out did not affect cell growth, whereas it increased SAM production by 29.3%, 55.6%, and 24.8% in G/Dsah, G/Dspe, and G/Dmsm, respectively, compared with the parental strain GS115/DS16. When l-Met addition was decreased from 0.10% to 0.06%, the SAM production increased by 26.4% and 28.9% in G/Dsah and G/Dmsm, respectively. [Conclusion] Therefore, the engineered P. pastoris strains can be utilized in industrial production of SAM in a cost-effective manner, and the strategy can also be employed for improving the production of other chemicals.

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苏慧颖,蒙世拯,赵欣欣,钱江潮,储炬,秦秀林. 阻断消耗途径提高毕赤酵母工程菌S-腺苷甲硫氨酸产量[J]. 微生物学通报, 2023, 50(2): 441-453

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历史
  • 收稿日期:2022-05-15
  • 最后修改日期:
  • 录用日期:2022-06-30
  • 在线发布日期: 2023-02-03
  • 出版日期: 2023-02-20