Efficient production of recombinant brazzein, a small, heat-stable, sweet-tasting protein of plant origin
Efficient production of recombinant brazzein, a small, heat-stable, sweet-tasting protein of plant origin
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DOI：
10.1006/abbi.2000.1725
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发表时间：
2000-04-15
影响因子：
3.9
通讯作者：
Markley, JL
中科院分区：
文献类型：
--
作者：
Assadi-Porter, FM;Aceti, DJ;Markley, JL
Brazzein是一种54氨基酸的甜味蛋白质，首先从西非的Pentadiplandra brazzeana Baillon的果实中分离出来。从水果中分离的植物甜蛋白，以重量计比蔗糖甜500倍（以每分子计甜9500倍）。来自水果的植物甜蛋白的次要成分，des-pGlu 1-brazzein具有53个氨基酸残基，并且具有亲本蛋白的两倍甜度。我们设计了一个des-pGlu 1- brazzein基因，该基因包含了在大肠杆菌中产生蛋白质的最佳密码子，从化学合成的基因产生的重组蛋白具有与从原始来源分离的brazzein相似的甜度，通过与具有设计的溴化氰切割位点的葡萄球菌核酸酶融合产生brazzein获得最佳产量。由于其强烈的甜味和在高pH值和温度下的稳定性，甜味蛋白是研究甜味特性所涉及的化学和结构要求的理想系统。这种高效的植物甜蛋白生产系统将有助于此类研究。(C)北京大学出版社.
Brazzein is: a 54-amino-acid sweet-tasting protein first isolated from the fruit of Pentadiplandra brazzeana Baillon found in West Africa. Brazzein, as isolated from the fruit, is 500 times sweeter than sucrose on a weight basis (9500 times sweeter on a per-molecule basis). A minor component of brazzein from fruit, des-pGlu1-brazzein has 53 amino acid residues and has twice the sweetness of the parent protein. We have designed a gene for des-pGlu1- brazzein that incorporates codons that are optimal for protein production in Escherichia coli, Production of brazzein from the chemically synthesized gene resulted in recombinant protein with sweetness similar to that of brazzein isolated from the original source, The best yields were achieved by producing brazzein as a fusion with staphylococcal nuclease with a designed cyanogen bromide cleavage site. Because of its intense sweetness and stability at high pH and temperature, brazzein is an ideal system for investigating the chemical and structural requirements involved in sweet-taste properties. This efficient protein production system for brazzein will facilitate such investigations. (C) 2000 Academic Press.