PMID- 35876873
OWN - NLM
STAT- MEDLINE
DCOM- 20220830
LR  - 20220830
IS  - 1432-0614 (Electronic)
IS  - 0175-7598 (Linking)
VI  - 106
IP  - 17
DP  - 2022 Sep
TI  - Recombinant production and characterization of L-glutaminase (glsA) as a 
      promiscuity therapeutic enzyme.
PG  - 5511-5524
LID - 10.1007/s00253-022-12058-y [doi]
AB  - Because of the therapeutical impacts of hydrolytic enzymes in different diseases, 
      in particular malignancies, we aimed to produce a recombinant putative 
      L-glutaminase (GLS A(SL-1)) from a recently characterized halo-thermotolerant 
      Bacillus sp. SL-1. For this purpose, the glsA gene was identified and efficiently 
      overexpressed in the Origami B (DE3) strain. The yield of the purified GLS 
      A(SL-1) was ~ 20 mg/L, indicating a significant expression of recombinant enzyme 
      in the Origami. The enzyme activity assay revealed a significant hydrolytic 
      effect of the recombinant GLS A(SL-1) on L-asparagine (Asn) (i.e., K(m) 39.8 muM, 
      k(cat) 19.9 S(-1)) with a minimal affinity for L-glutamine (Gln). The GLS A(SL-1) 
      significantly suppressed the growth of leukemic Jurkat cells through apoptosis 
      induction (47.5%) in the IC(50) dosage of the enzyme. The GLS A(SL-1) could also 
      change the Bax/Bcl2 expression ratio, indicating its apoptotic effect on cancer 
      cells. The in silico analysis was conducted to predict structural features 
      related to the histidine-tag exposure in the N- or C-terminal of the recombinant 
      GLS A(SL-1). In addition, molecular docking simulation for substrate specificity 
      revealed a greater binding affinity of Asn to the enzyme binding-site residues 
      than Gln, which was confirmed in experimental procedures as well. In conclusion, 
      the current study introduced a recombinant GLS A(SL-1) with unique functional and 
      structural features, highlighting its potential pharmaceutical and medical 
      importance. GLS A(SL-1) represents the first annotated enzyme from Bacillus with 
      prominent asparaginase activity, which can be considered for developing 
      alternative enzymes in therapeutic applications. KEY POINTS: * Hydrolytic enzymes 
      have critical applications in different types of human malignancies. * A 
      recombinant L-glutaminase (GLS A(SL-1)) was produced from halo-thermotolerant 
      Bacillus sp. SL-1. * GLS A(SL-1) displayed a marked hydrolytic activity on 
      L-asparagine compared to the L-glutamine. * GLS A(SL-1) with significant 
      substrate promiscuity may be an alternative for developing novel pharmaceuticals.
CI  - (c) 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, 
      part of Springer Nature.
FAU - Simay, Shayan
AU  - Simay S
AD  - Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz 
      University of Medical Sciences, Tabriz, Iran.
AD  - Department of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical 
      Sciences, Tabriz, Iran.
FAU - Akbarzadeh-Khiavi, Mostafa
AU  - Akbarzadeh-Khiavi M
AD  - Liver and Gastrointestinal Diseases Research Center, Tabriz University of Medical 
      Sciences, Tabriz, Iran.
FAU - Pourseif, Mohammad M
AU  - Pourseif MM
AD  - Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz 
      University of Medical Sciences, Tabriz, Iran.
FAU - Barar, Jaleh
AU  - Barar J
AD  - Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz 
      University of Medical Sciences, Tabriz, Iran.
AD  - Department of Pharmaceutics, Faculty of Pharmacy, Tabriz University of Medical 
      Sciences, Tabriz, Iran.
FAU - Safary, Azam
AU  - Safary A
AUID- ORCID: 0000-0002-8997-2003
AD  - Connective Tissue Diseases Research Center, Tabriz University of Medical 
      Sciences, Tabriz, Iran. azamsafary@yahoo.com.
FAU - Omidi, Yadollah
AU  - Omidi Y
AUID- ORCID: 0000-0003-0067-2475
AD  - Department of Pharmaceutical Sciences, College of Pharmacy, Nova Southeastern 
      University, Fort Lauderdale, FL, 33328, USA. yomidi@nova.edu.
LA  - eng
PT  - Journal Article
DEP - 20220725
PL  - Germany
TA  - Appl Microbiol Biotechnol
JT  - Applied microbiology and biotechnology
JID - 8406612
RN  - 0RH81L854J (Glutamine)
RN  - 7006-34-0 (Asparagine)
RN  - EC 3.5.1.1 (Asparaginase)
RN  - EC 3.5.1.2 (Glutaminase)
SB  - IM
MH  - Asparaginase
MH  - Asparagine
MH  - *Bacillus
MH  - Glutaminase
MH  - Glutamine
MH  - Humans
MH  - Molecular Docking Simulation
MH  - *Neoplasms
OTO - NOTNLM
OT  - Cloning
OT  - Halo-thermotolerant Bacillus
OT  - L-glutaminase
OT  - Molecular docking
OT  - Promiscuous function
OT  - Recombinant enzyme
EDAT- 2022/07/26 06:00
MHDA- 2022/08/31 06:00
CRDT- 2022/07/25 11:14
PHST- 2022/02/20 00:00 [received]
PHST- 2022/06/28 00:00 [accepted]
PHST- 2022/06/14 00:00 [revised]
PHST- 2022/07/26 06:00 [pubmed]
PHST- 2022/08/31 06:00 [medline]
PHST- 2022/07/25 11:14 [entrez]
AID - 10.1007/s00253-022-12058-y [pii]
AID - 10.1007/s00253-022-12058-y [doi]
PST - ppublish
SO  - Appl Microbiol Biotechnol. 2022 Sep;106(17):5511-5524. doi: 
      10.1007/s00253-022-12058-y. Epub 2022 Jul 25.