title: Recombinant expressions of sweet plant protein mabinlin II in Escherichia coli and food-grade Lactococcus lactis.
abstract: Sweet plant proteins, which are safe, natural, low-calorie sweeteners, may be suitable replacements for sugars in the food and beverage industries. Mabinlin II, a sweet plant protein, shows the most pronounced heat stability and acid resistance of any of the six known types of plant sweet proteins. However, mabinlin II is difficult to extract from the Capparis masaikai plant, which is itself becoming increasingly scarce. This limits the use of naturally acquired mabinlin II. In this study, recombinant mabinlin II proteins were expressed and purified in Escherichia coli and in food-grade Lactococcus lactis. Recombinant mabinlin II proteins MBL-BH (containing the B-chains of mabinlin II downstream fused with His-tag) and MBL-ABH (containing the A- and B-chains of mabinlin II downstream fused with His-tag) were expressed in E. coli in the form of inclusion bodies. They were then purified and renatured. The refolded MBL-BH was found to be 100 times sweeter than sucrose by weight, but it was not heat-stable. Refolded MBL-ABH was neither sweet nor heat-stable. Recombinant mabinlin II proteins were secreted and expressed intracellularly in food-grade L. lactis, in which the concentrated cell samples and culture medium samples were detected using enzyme-linked immunosorbent assay and Western blotting analysis with anti-mabinlin II polyclonal antibody. This study demonstrated that the single B chain of mabinlin II has a sweet taste. The recombinant mabinlin II proteins have been successfully expressed in food-grade L. lactis, which is a crucial step in the production of mabinlin II through microorganism expression systems.
authors: Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning, China, wenlianggu@gmail.com.
authors: Wenliang
authors: W
authors: Gu
authors: Qiyu
authors: Q
authors: Xia
authors: Jing
authors: J
authors: Yao
authors: Shaoping
authors: S
authors: Fu
authors: Jianchun
authors: J
authors: Guo
authors: Xinwen
authors: X
authors: Hu
mesh_terms: D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005516:Food Microbiology; D015870:Gene Expression; D013294:Lactococcus lactis / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D013549:Sweetening Agents / Q000378:metabolism*
chemical_list: D010940:Plant Proteins; D013549:Sweetening Agents; C083584:mabinlin protein, Capparis masaikai
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