PMID- 34622632
OWN - NLM
STAT- MEDLINE
DCOM- 20211011
LR  - 20220531
IS  - 1872-2075 (Electronic)
IS  - 1000-3061 (Linking)
VI  - 37
IP  - 9
DP  - 2021 Sep 25
TI  - [Molecular modification and highly efficient expression of L-asparaginase from 
      Rhizomucor miehei].
PG  - 3242-3252
LID - 10.13345/j.cjb.200759 [doi]
AB  - L-asparaginase hydrolyzes L-asparagine to produce L-aspartic acid and ammonia. It 
      is widely distributed in microorganisms, plants and serum of some rodents, and 
      has important applications in the pharmaceutical and food industries. However, 
      the poor thermal stability, low catalytic efficiency and low yield hampered the 
      further application of L-asparaginase. In this paper, rational design and 5' 
      untranslated region (5'UTR) design strategies were used to increase the specific 
      enzyme activity and protein expression of L-asparaginase derived from Rhizomucor 
      miehei (RmAsnase). The results showed that among the six mutants constructed 
      through homology modeling combined with sequence alignment, the specific enzyme 
      activity of the mutant A344E was 1.5 times higher than the wild type. 
      Subsequently, a food-safe strain Bacillus subtilis 168/pMA5-A344E was 
      constructed, and the UTR strategy was used for the construction of recombinant 
      strain B. subtilis 168/pMA5 UTR-A344E. The enzyme activity of B. subtilis 
      168/pMA5 UTR-A344E was 7.2 times higher than that of B. subtilis 168/pMA5-A344E. 
      The recombinant strain B. subtilis 168/pMA5 UTR-A344E was scaled up in 5 L 
      fermenter, and the final yield of L-asparaginase was 489.1 U/mL, showing great 
      potential for industrial application.
FAU - Zhu, Manchi
AU  - Zhu M
AD  - Key Laboratory of Industrial Biotechnology of Ministry of Education, School of 
      Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
FAU - Zhang, Xian
AU  - Zhang X
AD  - Key Laboratory of Industrial Biotechnology of Ministry of Education, School of 
      Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
FAU - Wang, Zhi
AU  - Wang Z
AD  - Key Laboratory of Industrial Biotechnology of Ministry of Education, School of 
      Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
FAU - Lin, Wenxuan
AU  - Lin W
AD  - Key Laboratory of Industrial Biotechnology of Ministry of Education, School of 
      Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
FAU - Xu, Meijuan
AU  - Xu M
AD  - Key Laboratory of Industrial Biotechnology of Ministry of Education, School of 
      Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
FAU - Yang, Taowei
AU  - Yang T
AD  - Key Laboratory of Industrial Biotechnology of Ministry of Education, School of 
      Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
FAU - Shao, Minglong
AU  - Shao M
AD  - Key Laboratory of Industrial Biotechnology of Ministry of Education, School of 
      Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
FAU - Rao, Zhiming
AU  - Rao Z
AD  - Key Laboratory of Industrial Biotechnology of Ministry of Education, School of 
      Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
LA  - chi
PT  - Journal Article
PL  - China
TA  - Sheng Wu Gong Cheng Xue Bao
JT  - Sheng wu gong cheng xue bao = Chinese journal of biotechnology
JID - 9426463
RN  - EC 3.5.1.1 (Asparaginase)
RN  - Rhizomucor miehei
SB  - IM
MH  - *Asparaginase/biosynthesis/genetics
MH  - Bacillus subtilis/genetics
MH  - Industrial Microbiology
MH  - Protein Engineering
MH  - *Rhizomucor/enzymology
MH  - Sequence Alignment
OTO - NOTNLM
OT  - L-asparaginase
OT  - UTR strategy
OT  - site-directed mutation
OT  - specific enzyme activity
EDAT- 2021/10/09 06:00
MHDA- 2021/10/12 06:00
CRDT- 2021/10/08 07:38
PHST- 2021/10/08 07:38 [entrez]
PHST- 2021/10/09 06:00 [pubmed]
PHST- 2021/10/12 06:00 [medline]
AID - 10.13345/j.cjb.200759 [doi]
PST - ppublish
SO  - Sheng Wu Gong Cheng Xue Bao. 2021 Sep 25;37(9):3242-3252. doi: 
      10.13345/j.cjb.200759.