Characterization of a new microbial Pictet-Spenglerase NscbB affording the β-carboline skeletons from Nocardiopsis synnemataformans DSM 44143
Characterization of a new microbial Pictet-Spenglerase NscbB affording the β-carboline skeletons from Nocardiopsis synnemataformans DSM 44143
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DOI：
10.1016/j.jbiotec.2018.07.007
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发表时间：
2018-09-10
影响因子：
4.1
通讯作者：
Xie, Yunchang
中科院分区：
文献类型：
--
作者：
Chen, Qi;Zhang, Shaofei;Xie, Yunchang
β-咔啉（beta-C）生物碱生物合成的特征是由Pictet-Spengler酶（PSase）催化的酶促Pictet-Spengler（PS）反应。NscbB是一种罕见的微生物PSase，通过与其充分表征的对应物McbB的同源比对，从Nocardiopsis syndrome aformans DSM 44143（肾移植患者衍生的）中的隐蔽β-咔啉生物碱生物合成基因簇（BGC）中发现。生化分析表明，NscbB在体外可催化L-色氨酸（K-M = 89.64 +/- 8.69 μ M）和甲基乙二醛（K-M = 147.70 +/- 16.38 μ M）在无辅因子的情况下形成两个β C骨架1-乙酰基-3-羧基-β-咔啉和1-乙酰基-β-咔啉。此外，nscbB在E. coli BL 21（DE 3）中，NscbB在16 h内可高效生产2种β-C骨架，1-乙酰基-3-羧基-β-咔啉（5.5 mg/L）和1-乙酰基-β-咔啉（3.1 mg/L）。这些结果表明，NscbB是一种新的和实用的微生物PSase，这是未来的生物活性β C生物碱的开发和发展。
The enzymatic Pictet-Spengler (PS) reaction, catalyzed by Pictet-Spenglerase (PSase), is the feature of beta-carboline (beta C) alkaloid biosynthesis. NscbB is a rare microbial PSase discovered from a cryptic beta-carboline alkaloid biosynthetic gene cluster (BGC) in the Nocardiopsis synnemataformans DSM 44143 (kidney transplant patient derived) by homologous alignment with its well-characterized counterpart McbB. The biochemical analysis showed that NscbB could catalyze L-tryptophan (K-M = 89.64 +/- 8.69 mu M) and methylglyoxal (K-M = 147.70 +/- 16.38 mu M), without cofactors, to form the two beta C skeletons 1-acetyl-3-carboxy-beta-carboline and 1-acetyl-beta-carboline in vitro. Additionally, the heterologous expression of nscbB in E. coli BL21 (DE3) revealed the efficient bioproductivity of NscbB to bioproduce two beta C skeletons, 1-acetyl-3-carboxy-beta-carboline (5.5 mg/L) and 1-acetyl-beta-carboline (3.1 mg/L), within 16 h fermentation. These results demonstrate NscbB is a novel and practical microbial PSase, which is useful for future bioactive beta C alkaloid exploitation and development.