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通过精准发酵利用重组酵母分泌生产含血红素的植物珠蛋白

Secretory Production of Plant Heme-Containing Globins by Recombinant Yeast via Precision Fermentation.

作者信息

Bae Ha-Neul, Kim Geun-Hyung, Seo Seung-Oh

机构信息

Department of Food Science and Biotechnology, Seoul National University of Science and Technology, Seoul 01811, Republic of Korea.

Research Institute of Food and Biotechnology, Seoul National University of Science and Technology, Seoul 01811, Republic of Korea.

出版信息

Foods. 2025 Apr 20;14(8):1422. doi: 10.3390/foods14081422.

Abstract

Leghemoglobin (LegHb) is a plant-derived heme-containing globin found in the root nodules of legumes like soybean that can be used as a food additive for red color and meaty flavor as a plant-based meat alternative. However, conventional extraction methods face challenges of low yield and high costs. To address this issue, precision fermentation with recombinant microorganisms has been applied for the sustainable large-scale production of plant leghemoglobins. This study attempted the production of plant legHbs using recombinant yeast strains, and . The plant legHb genes were identified from the genome of legumes such as soybean, chickpea, mung bean and overexpressed in yeast via extracellular secretion by the signal peptide and inducible promoters. Subsequently, hemin as a heme provider was added to the fermentation, resulting in increased levels of plant legHbs. In , gmaLegHb expression reached up to 398.1 mg/L, while in , gmaLegHb showed the highest production level, reaching up to 1652.7 mg/L. The secretory production of plant legHbs was further enhanced by replacing the signal peptide in the recombinant yeast. The secreted plant legHbs were purified by His-Tag from a culture supernatant or concentrated via precipitation using ammonium sulfate. These results suggest that the production of plant legHbs is significantly influenced by hemin and signal peptide. This study successfully demonstrates the production of the various plant legHbs other than soy legHb that can be used as natural colors and flavors for plant-based meat alternatives.

摘要

豆血红蛋白(LegHb)是一种源自植物的含血红素球蛋白,存在于大豆等豆类植物的根瘤中,可作为一种植物性肉类替代品的红色素和肉味食品添加剂。然而,传统的提取方法面临着产量低和成本高的挑战。为了解决这个问题,利用重组微生物进行精准发酵已被应用于植物豆血红蛋白的可持续大规模生产。本研究尝试使用重组酵母菌株生产植物豆血红蛋白。从大豆、鹰嘴豆、绿豆等豆类植物的基因组中鉴定出植物豆血红蛋白基因,并通过信号肽和诱导型启动子在酵母中进行细胞外分泌过表达。随后,在发酵过程中添加血红素作为血红素供体,从而提高了植物豆血红蛋白的水平。在[具体实验条件1]中,gmaLegHb的表达量达到398.1毫克/升,而在[具体实验条件2]中,gmaLegHb的产量最高,达到1652.7毫克/升。通过替换重组酵母中的信号肽,进一步提高了植物豆血红蛋白的分泌产量。分泌的植物豆血红蛋白通过His标签从培养上清液中纯化,或通过硫酸铵沉淀进行浓缩。这些结果表明,血红素和信号肽对植物豆血红蛋白的生产有显著影响。本研究成功证明了除大豆豆血红蛋白外,其他多种植物豆血红蛋白的生产,这些豆血红蛋白可作为植物性肉类替代品的天然色素和风味剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/91c9/12026525/149c949fc011/foods-14-01422-g001.jpg

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