Wang Yanhong, Wang Xinhui, Jiang Peng, Dai Lingyan, Hu Yijia, Pan Bailing, Li Yueyue, Zhang Jingyu, Zhang Ruoyu, Zhan Shihan, Li Zhijiang
Heilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argro-Waste in Cold Region, Department of Bioengineering, College of Science and Biotechnology Heilongjiang Bayi Agricultural University Daqing Heilongjiang China.
Department of Food Science and Engineering, College of Food Science Heilongjiang Bayi Agricultural University Daqing Heilongjiang China.
Food Sci Nutr. 2024 Mar 18;12(6):4435-4442. doi: 10.1002/fsn3.4109. eCollection 2024 Jun.
4-Hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) is a flavor compound widely found in natural products and is used in food as a flavor-enhancing agent. Quinone oxidoreductase (QOR) was verified as a key enzyme to synthesize HDMF in strawberry, while its impact on HDMF production by was still unknown. The gene was cloned and overexpressed in , and its impact on HDMF production by was then further analyzed. At the same time, it is expected to obtain engineered strains of with high HDMF production. The results showed that the engineered strains of exhibit different levels of gene expression and HDMF production; among them, the QOR6 strain exhibiting the highest gene expression level and HDMF production was named as . The HDMF production of the strain was significantly higher than that of wild-type at 3 and 5 days of culture, with 1.41-fold and 1.08-fold increases, respectively. At 3 days of fermentation, the highest HDMF yield of strain was obtained (2.75 mg/L), 2 days ahead of the reported highest HDMF production by . At 3, 5, and 7 days, gene expression was 4.8-fold, 3.3-fold, and 5.6-fold higher in the strain than in the wild-type , respectively. Therefore, overexpression of the gene facilitates HDMF synthesis. The genetic stability of the 0-20 generation strain was stable, and there was no significant difference in colony shape, expression, or HDMF production compared to the wild type. In this study, the genetic engineering strain was used to improve HDMF production. This research has laid the groundwork for further industrial production of HDMF via microbial synthesis.
4-羟基-2,5-二甲基-3(2H)-呋喃酮（HDMF）是一种广泛存在于天然产物中的风味化合物，在食品中用作增味剂。醌氧化还原酶（QOR）被证实是草莓中合成HDMF的关键酶，但其对[具体对象]HDMF产量的影响尚不清楚。[具体基因]基因被克隆并在[具体对象]中过表达，然后进一步分析其对[具体对象]HDMF产量的影响。同时，期望获得具有高HDMF产量的[具体对象]工程菌株。结果表明，[具体对象]工程菌株表现出不同水平的[具体基因]表达和HDMF产量；其中，基因表达水平和HDMF产量最高的QOR6菌株被命名为[具体名称]。[具体名称]菌株在培养3天和5天时的HDMF产量显著高于野生型[具体对象]，分别增加了1.41倍和1.08倍。在发酵3天时，[具体名称]菌株获得了最高的HDMF产量（2.75mg/L），比报道的[具体对象]最高HDMF产量提前了2天。在3天、5天和7天时，[具体名称]菌株中的[具体基因]表达分别比野生型[具体对象]高4.8倍、3.3倍和5.6倍。因此，[具体基因]的过表达促进了HDMF的合成。0-20代[具体名称]菌株的遗传稳定性良好，与野生型相比，菌落形态、[具体基因]表达或HDMF产量均无显著差异。在本研究中，利用基因工程[具体名称]菌株提高了HDMF产量。该研究为通过微生物合成进一步工业化生产HDMF奠定了基础。
