Tryptophan production by catalysis of a putative tryptophan synthase protein

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS
Lulu Cao, Jiaqi Zhang, Jia Chen, Mei Li, Hao Chen, Chongju Wang, Chunjie Gong
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Abstract

Essential amino acid, tryptophan which intake from food plays a critical role in numerous metabolic functions, exhibiting extensive biological functions and applications. Tryptophan is beneficial for the food sector by enhancing nutritional content and promoting the development of functional foods. A putative gene encoding tryptophan synthase was the first identified in Sphingobacterium soilsilvae Em02, a cellulosic bacterium making it inherently more environmentally friendly. The gene was cloned and expressed in exogenous host Escherichia coli, to elucidate its function. The recombinant tryptophan synthase with a molecular weight 42 KDa was expressed in soluble component. The enzymatic activity to tryptophan synthase in vivo was assessed using indole and L-serine and purified tryptophan synthase. The optimum enzymatic activity for tryptophan synthase was recorded at 50 ºC and pH 7.0, which was improved in the presence of metal ions Mg2+, Sr2+ and Mn2+, whereas Cu2+, Zn2+ and Co2+ proved to be inhibitory. Using site-directed mutagenesis, the consensus pattern HK-S-[GGGSN]-E-S in the tryptophan synthase was demonstrated with K100Q, S202A, G246A, E361A and S385A as the active sites. Tryptophan synthase has been demonstrated to possess the defining characteristics of the β-subunits. The tryptophan synthase may eventually be useful for tryptophan production on a larger scale. Its diverse applications highlight the potential for improving both the quality and health benefits of food products, making it an essential component in advancing food science and technology.

Abstract Image

通过推测的色氨酸合成酶蛋白的催化作用产生色氨酸。
从食物中摄取的必需氨基酸色氨酸在许多新陈代谢功能中发挥着关键作用,具有广泛的生物功能和应用。色氨酸可提高营养成分,促进功能性食品的开发,对食品行业大有裨益。首次在纤维素细菌 Sphingobacterium soilsilvae Em02 中发现了一个编码色氨酸合成酶的推测基因,该基因使纤维素细菌更加环保。该基因被克隆并在外源宿主大肠杆菌中表达,以阐明其功能。重组色氨酸合成酶的分子量为 42 KDa,以可溶性成分表达。使用吲哚和 L-丝氨酸以及纯化的色氨酸合成酶评估了色氨酸合成酶在体内的酶活性。色氨酸合成酶的最佳酶活性是在 50 ºC 和 pH 值为 7.0 时记录的,在金属离子 Mg2+、Sr2+ 和 Mn2+ 的存在下,酶活性得到提高,而 Cu2+、Zn2+ 和 Co2+ 被证明具有抑制作用。通过定点突变,色氨酸合成酶中的共识模式 HK-S-[GGGSN]-E-S被证实,K100Q、S202A、G246A、E361A 和 S385A 为活性位点。色氨酸合成酶已被证明具有 β 亚基的定义特征。色氨酸合成酶最终可能会用于更大规模的色氨酸生产。色氨酸合成酶的多样化应用凸显了其在提高食品质量和健康益处方面的潜力,使其成为推动食品科学与技术发展的重要组成部分。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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