瑞士乳杆菌CNRZ32中一种与细胞壁相关的蛋白酶的遗传特征分析
Genetic characterization of a cell envelope-associated proteinase from Lactobacillus helveticus CNRZ32.
作者信息
Pederson J A, Mileski G J, Weimer B C, Steele J L
机构信息
Department of Food Science, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
出版信息
J Bacteriol. 1999 Aug;181(15):4592-7. doi: 10.1128/JB.181.15.4592-4597.1999.
Abstract
A cell envelope-associated proteinase gene (prtH) was identified in Lactobacillus helveticus CNRZ32. The prtH gene encodes a protein of 1,849 amino acids and with a predicted molecular mass of 204 kDa. The deduced amino acid sequence of the prtH product has significant identity (45%) to that of the lactococcal PrtP proteinases. Southern blot analysis indicates that prtH is not broadly distributed within L. helveticus. A prtH deletion mutant of CNRZ32 was constructed to evaluate the physiological role of PrtH. PrtH is not required for rapid growth or fast acid production in milk by CNRZ32. Cell surface proteinase activity and specificity were determined by hydrolysis of alpha(s1)-casein fragment 1-23 by whole cells. A comparison of CNRZ32 and its prtH deletion mutant indicates that CNRZ32 has at least two cell surface proteinases that differ in substrate specificity.
摘要
在瑞士乳杆菌CNRZ32中鉴定出一种与细胞包膜相关的蛋白酶基因（prtH）。prtH基因编码一种含有1849个氨基酸的蛋白质，预测分子量为204 kDa。prtH产物推导的氨基酸序列与乳球菌PrtP蛋白酶的氨基酸序列具有显著的同一性（45%）。Southern杂交分析表明，prtH在瑞士乳杆菌中分布不广泛。构建了CNRZ32的prtH缺失突变体以评估PrtH的生理作用。CNRZ32在牛奶中快速生长或快速产酸不需要PrtH。通过全细胞对α(s1)-酪蛋白片段1-23的水解来测定细胞表面蛋白酶的活性和特异性。对CNRZ32及其prtH缺失突变体的比较表明，CNRZ32至少有两种细胞表面蛋白酶，它们的底物特异性不同。
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