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Penicillin G acylase production by Mucor griseocyanus and the partial genetic analysis of its pga gene.
International Microbiology ( IF 2.3 ) Pub Date : 2020-07-23 , DOI: 10.1007/s10123-020-00137-x
Juan C Cano-Cabrera 1 , Lissethe Palomo-Ligas 2 , Adriana C Flores-Gallegos 2 , José L Martínez-Hernández 1 , Raúl Rodríguez-Herrera 2
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Penicillin acylases (penicillin amidohydrolase, EC 3.5.1.11) are a group of enzymes with many applications within the pharmaceutical industry, and one of them is the production of semi-synthetic beta-lactam antibiotics. This enzyme is mainly produced by bacteria but also by some fungi. In the present study, the filamentous fungus Mucor griseocyanus was used to produce penicillin acylase enzyme (PGA). Its ability to express PGA enzyme in submerged fermentation process was assessed, finding that this fungal strain produces the biocatalyst of interest in an extracellular way at a level of 570 IU/L at 72 h of fermentation; in this case, a saline media using lactose as carbon source and penicillin G as inducer was employed. In addition, a DNA fragment (859 bp) of the pga from a pure Mucor griseocyanus strain was amplified, sequenced, and analyzed in silico. The partial sequence of pga identified in the fungi showed high identity percentage with penicillin G acylase sequences deposited in NCBI through BLAST, especially with the β subunit of PGA from the Alcaligenes faecalis bacterium¸ which is a region involved in the catalytic function of this protein. Besides, the identification of domains in the penicillin G acylase sequence of Mucor griseocyanus showed three conserved regions of this protein. The bioinformatic results support the identity of the gen as penicillin G acylase. This is the first report that involves sequencing and in silico analysis of Mucor griseocyanus strain gene encoding PGA.



中文翻译:

灰蓝毛霉生产青霉素 G 酰化酶及其 pga 基因的部分遗传分析。

青霉素酰化酶(penicillin amidohydrolase,EC 3.5.1.11)是一组在制药工业中有许多应用的酶,其中之一是生产半合成β-内酰胺抗生素。这种酶主要由细菌产生,但也由一些真菌产生。在本研究中,丝状真菌Mucor griseocyanus用于生产青霉素酰化酶 (PGA)。对其在深层发酵过程中表达 PGA 酶的能力进行了评估,发现该真菌菌株在发酵 72 小时时以细胞外方式以 570 IU/L 的水平产生感兴趣的生物催化剂;在这种情况下,采用以乳糖为碳源和青霉素 G 为诱导剂的盐水培养基。此外,pga的 DNA 片段 (859 bp)来自纯灰蓝毛霉菌株被扩增、测序和计算机分析的部分序列PGA在真菌识别的显示高同一性百分比用青霉素G酰基转移酶通过BLAST沉积在NCBI序列,尤其是与来自PGA的β亚基粪产碱杆菌细菌¸这是参与该蛋白质的催化功能的区域。此外,灰蓝毛霉青霉素 G 酰化酶序列中的域的鉴定显示该蛋白质的三个保守区域。生物信息学结果支持该基因为青霉素 G 酰基转移酶。这是第一份涉及测序和计算机分析的报告灰蓝毛霉菌株基因编码 PGA。

更新日期:2020-07-23
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