PMID- 27516945
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20160812
LR  - 20231111
IS  - 2193-1801 (Print)
IS  - 2193-1801 (Electronic)
IS  - 2193-1801 (Linking)
VI  - 5
IP  - 1
DP  - 2016
TI  - Efficient 9alpha-hydroxy-4-androstene-3,17-dione production by engineered Bacillus 
      subtilis co-expressing Mycobacterium neoaurum 3-ketosteroid 9alpha-hydroxylase and B. 
      subtilis glucose 1-dehydrogenase with NADH regeneration.
PG  - 1207
LID - 10.1186/s40064-016-2871-4 [doi]
LID - 1207
AB  - 3-Ketosteroid 9alpha-hydroxylase (KSH, consisting of KshA and KshB), a key enzyme in 
      steroid metabolism, can catalyze the transformation of 4-androstene-3,17-dione 
      (AD) to 9alpha-hydroxy-4-androstene-3,17-dione (9OHAD) with NADH as coenzyme. In this 
      work, KSH from Mycobacterium neoaurum JC-12 was successfully cloned and 
      overexpressed in Bacillus subtilis 168. The expression and purification of KSH 
      was analyzed by SDS-PAGE and KSH activity assay. Preliminary characterization of 
      KSH was performed using purified KshA and KshB. The results showed that KSH was 
      very unstable, and its activity was inhibited by most metal ions, especially 
      Zn(2+). The whole-cells of recombinant B. subtilis, co-expression of KSH and 
      glucose 1-dehydrogenase (GDH), were used as biocatalyst to convert AD to 9OHAD. 
      The biocatalyst, in which the intracellular NADH was regenerated, efficiently 
      catalyzed the bioconversion of AD to 9OHAD with a conversion rate of 90.4 % and 
      productivity of 0.45 g (L h)(-1), respectively. This work proposed a strategy for 
      efficiently producing 9OHAD by using B. subtilis as a promising whole-cell 
      biocatalyst host and co-expressing KSH and GDH to construct a NADH regeneration 
      system.
FAU - Zhang, Xian
AU  - Zhang X
AD  - The Key Laboratory of Industrial Biotechnology of Ministry of Education, School 
      of Biotechnology, Jiangnan University, Wuxi, 214122 Jiangsu People's Republic of 
      China.
FAU - Rao, Zhiming
AU  - Rao Z
AD  - The Key Laboratory of Industrial Biotechnology of Ministry of Education, School 
      of Biotechnology, Jiangnan University, Wuxi, 214122 Jiangsu People's Republic of 
      China.
FAU - Zhang, Lele
AU  - Zhang L
AD  - Jiangnan University (Rugao) Food Biotechnology Research Institute, Jiangsu 
      Industrial Technology Research Institute, Rugao, 226500 Jiangsu People's Republic 
      of China.
FAU - Xu, Meijuan
AU  - Xu M
AD  - The Key Laboratory of Industrial Biotechnology of Ministry of Education, School 
      of Biotechnology, Jiangnan University, Wuxi, 214122 Jiangsu People's Republic of 
      China.
FAU - Yang, Taowei
AU  - Yang T
AD  - The Key Laboratory of Industrial Biotechnology of Ministry of Education, School 
      of Biotechnology, Jiangnan University, Wuxi, 214122 Jiangsu People's Republic of 
      China.
LA  - eng
GR  - N01AA21004/HD/NICHD NIH HHS/United States
GR  - R25 AA021304/AA/NIAAA NIH HHS/United States
PT  - Journal Article
DEP - 20160729
PL  - Switzerland
TA  - Springerplus
JT  - SpringerPlus
JID - 101597967
PMC - PMC4967059
OTO - NOTNLM
OT  - 3-Ketosteroid 9alpha-hydroxylase
OT  - 9alpha-Hydroxy-4-androstene-3,17-dione
OT  - Bacillus subtilis
OT  - Mycobacterium neoaurum
EDAT- 2016/08/16 06:00
MHDA- 2016/08/16 06:01
PMCR- 2016/07/29
CRDT- 2016/08/13 06:00
PHST- 2016/05/13 00:00 [received]
PHST- 2016/07/19 00:00 [accepted]
PHST- 2016/08/13 06:00 [entrez]
PHST- 2016/08/16 06:00 [pubmed]
PHST- 2016/08/16 06:01 [medline]
PHST- 2016/07/29 00:00 [pmc-release]
AID - 2871 [pii]
AID - 10.1186/s40064-016-2871-4 [doi]
PST - epublish
SO  - Springerplus. 2016 Jul 29;5(1):1207. doi: 10.1186/s40064-016-2871-4. eCollection 
      2016.