Molecular Cloning of Microbial Transglutaminase from Streptoverticillium ladakanum BCRC 12422

Streptoverticillium ladakanum BCRC 12422轉麩胺醯胺酶之基因選殖

曾鑫順(Shinn-Shuenn Tzeng) ； 殷儷容(Li-Jung Yin) ； 陳俊宏(Gen-Hung Chen) ； 江善宗(Shann-Tzong Jiang)

《臺灣水產學會刊》 32卷1期 (2005/03) Pp. 53-67

https://doi.org/10.29822/JFST.200503.0005

摘要

轉麩胺醯胺酶對於蛋白質功能性具有潛在的修飾作用。Streptoverticillium ladakanum生產之轉麩胺醯胺酶經由質譜儀測得之分子量為38210，而其胺端序列經測定為RAPDSDERVTPP。本研究成功選殖出含有轉麩胺醯胺酶的核苷酸片段，該序列由1497鹼基對所組成，可推導出一個具有410個胺基酸的完整蛋白質，包括了可能為訊息序列的76個胺基酸，以及構成活性酵素的334個胺基酸，且該胺基酸序列之分子量計算值近似於質譜儀上的測定值。此外，該酵素胺基酸序列與放射線菌類之微生物轉麩胺醯胺酶相似度高達80%以上，而活性酵素序列中的單－Cys更可能在酵素的催化作用中扮演極為重要的角色。

關鍵字

Streptoverticillium ladakanum ； 轉麩胺醯胺酶 ； 基因選殖 ； 核苷酸序列

並列摘要

Transglutaminase (TGase) has potential application in the modification of protein functionalities. The molecular mass of microbial TGase (MTGase) from Streptoverticillium ladakanum was 38210 as determined by mass spectrometer. Its N-terminal dodecapeptide determined by automated Edman degradation was RAPDSDERVTPP. A genetic sequence of 1497 base pairs nucleotides encoding MTGase, which could deduce a polypeptide chain containing 410 amino acid residues, was cloned and expressed in E. coil AD494 (DE3) after obtaining the full-length sequence. The expressed MTGase was composed of the prepro-domain and mature form enzyme, which had 76 and 334 amino acid residues, respectively. The calculated molecular mass of mature MTGase coincided with its mass value. Comparison of amino acid sequence suggested that the enzyme was homologous to actinomycete MTGases with ＞80% identity and the sole Cys in mature enzyme might play crucial roles in catalytic activity.

並列關鍵字

Streptoverticillium ladakanum ； microbial transglutaminase ； molecular cloning, nucleotide sequence