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        Extracellular Proteome Profiling of Bacillus pumilus SCU11 Producing Alkaline Protease for Dehairing<sup>s</sup>

        ( Chao Wang ),( Shiqiang Yu ),( Ting Song ),( Tingting He ),( Huanhuan Shao ),( Haiyan Wang ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.11

        Bacillus pumilus is one of the most characterized microorganisms that are used for high-level production of select industrial enzymes. A novel B. pumilus SCU11 strain possessing high alkaline protease activity was obtained in our previous work. The culture supernatant of this strain showed efficient dehairing capability with minimal collagen damage, indicating promising potential applications in the leather industry. In this study, the strain`s extracellular proteome was identified by LC-MS/MS-based shotgun proteomic analysis, and their related secretory pathways were characterized by BLAST searches. A total of 513 proteins, including 100 actual secreted and 413 intracellular proteins, were detected in the extracellular proteome. The functions of these secreted proteins were elucidated and four complete secretory systems (Sec, Tat, Com, and ABC transporter) were proposed for B. pumilus. These data provide B. pumilus a comprehensive extracellular proteome profile, which is a valuable theoretical and applicative basis for future genetic modifications and development of industrial enzymes.

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        Degradation Characteristics and Metabolic Pathway of 17β-estradiol (E2) by Rhodococcus sp. DS201

        Qingmiao Yu,Ping Wang,Dongbo Liu,Ruixia Gao,Huanhuan Shao,Hongyan Zhao,Zhe Ma,Dan Wang,Hongliang Huo 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.6

        In this study an E2-degrading bacterium was isolated from the activated sludge of a municipal treatment plant that treats the waste from a contraceptive medicineprocessing factory in Beijing, China. Using the observed morphological and physiological features of the bacterium and 16S rRNA sequence analysis, this bacterial strain was identified as Rhodococcus sp. DS201. Using single-factor experiments and orthogonal tests, it was demonstrated that, when strain DS210 bacteria were inoculated into MM medium at an initial concentration of 1 mg/L with an initial pH of 7 and an inoculum amount of 1%, complete degradation of E2 by this strain was achieved within 3 days at 30oC. After strain DS201 had degraded the E2, several E2 metabolites were detected in the culture extracts using high-performance liquid chromatography (HPLC); they were then further identified using HPLC with tandem mass spectrometry (LC-MS/MS). Mass spectrum analysis of the E2 degradation identified the following products: pent- 4-enoic acid; 2-ethyl-3-hydroxy-6-methylcyclohexane-1- carboxylic acid; 3-(7a-methyl-1,5-dioxooctahydro-1H-inden- 4-yl) propanoic acid; and 5-hydroxy-4-(3-hydroxypropyl)- 7a-methyloctahydro-1H-inden-1-one. These products have not previously been reported as parts of a mechanism for microbial E2 degradation and were suspected to be new metabolite products. Therefore, the E2 degradation pathway by strain DS201 is proposed herein.

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