PMID- 37815749
OWN - NLM
STAT- Publisher
LR  - 20231010
IS  - 1559-0305 (Electronic)
IS  - 1073-6085 (Linking)
DP  - 2023 Oct 10
TI  - Identification of Cutinolytic Esterase from Microplastic-Associated Microbiota 
      Using Functional Metagenomics and Its Plastic Degrading Potential.
LID - 10.1007/s12033-023-00916-7 [doi]
AB  - Plastic pollution has threatened biodiversity and human health by shrinking 
      habitats, reducing food quality, and limiting the activities of organisms. 
      Therefore, global interest in discovering novel enzymes capable of degrading 
      plastics has increased considerably. Within this context, the functional 
      metagenomic approach, which allows for unlocking the functional potential of 
      uncultivable microbial biodiversity, was used to discover a plastic-degrading 
      enzyme. First, metagenomic libraries derived from microplastic-associated 
      microbiota were screened for esterases capable of degrading both tributyrin and 
      polycaprolactone. Clone KAD01 produced esterase highly active against 
      p-nitrophenyl esters (C2-C16). The gene corresponding to the enzyme activity 
      showed moderate identity (</= 55.94%) to any known esterases/cutinases. The gene 
      was extracellularly expressed with a 6x histidine tag in E. coli BL21(DE3), 
      extracellularly. Titer of the enzyme (CEstKAD01) was raised from 21.32 to 
      35.17 U/mL by the statistical optimization of expression conditions and media 
      components. CEstKAD01 was most active at pH 7.0 and 30  degrees C. It was noteworthy 
      stable over a wide pH (6.0-10.0) and temperature (20-50  degrees C). The enzyme was 
      active and stable in elevated NaCl concentrations up to 12% (w/v). Pre-incubation 
      of CEstKAD01 with Mg(2+), Mn(2+), and Ca(2+) increased the enzyme activity. 
      CEstKAD01 displayed an excellent tolerance against various chemicals and 
      solvents. It was determined that 1 mg of the enzyme caused the release of 
      5.39 +/- 0.18 mM fatty acids from 1 g apple cutin in 120 min. K(m) and V(max) 
      values of CEstKAD01 against p-nitrophenyl butyrate were calculated to be 1.48 mM 
      and 20.37 micromol/min, respectively. The enzyme caused 6.94 +/- 0.55, 8.71 +/- 0.56, 
      7.47 +/- 0.47, and 9.22 +/- 0.18% of weight loss in polystyrene, high-density 
      polyethylene, low-density polyethylene, and polyvinyl chloride after 30-day 
      incubation. The scanning electron microscopy (SEM) analysis indicated the 
      formation of holes and pits on the plastic surfaces supporting the degradation. 
      In addition, the change in chemical structure in plastics treated with the enzyme 
      was determined by Fourier Transform Infrared Spectroscopy (FTIR) analysis. 
      Finally, the degradation products were found to have no genotoxic potential. To 
      our knowledge, no cutinolytic esterase with the potential to degrade polystyrene 
      (PS), high-density polyethylene (HDPE), low-density polyethylene (LDPE), and 
      polyvinyl chloride (PVC) has been identified from metagenomes derived from 
      microplastic-associated microbiota.
CI  - (c) 2023. The Author(s), under exclusive licence to Springer Science+Business 
      Media, LLC, part of Springer Nature.
FAU - Adiguzel, Ali Osman
AU  - Adiguzel AO
AUID- ORCID: 0000-0002-5602-5886
AD  - Department of Molecular Biology and Genetics, Faculty of Science, Ondokuz Mayis 
      University, Samsun, 55000, Turkey. adiguzel.ali.osman@gmail.com.
FAU - Sen, Fatma
AU  - Sen F
AD  - Department of Molecular Biology and Genetics, Faculty of Science, Ondokuz Mayis 
      University, Samsun, 55000, Turkey.
FAU - Konen-Adiguzel, Serpil
AU  - Konen-Adiguzel S
AD  - Department of Biology, Faculty of Science, Mersin University, Mersin, Turkey.
FAU - Kideys, Ahmet Erkan
AU  - Kideys AE
AD  - Department of Marine Biology and Fisheries, Institute of Marine Sciences, Middle 
      East Technical University, Mersin, Turkey.
FAU - Karahan, Arzu
AU  - Karahan A
AD  - Department of Marine Biology and Fisheries, Institute of Marine Sciences, Middle 
      East Technical University, Mersin, Turkey.
FAU - Doruk, Tugrul
AU  - Doruk T
AD  - Department of Molecular Biology and Genetics, Faculty of Science, Ondokuz Mayis 
      University, Samsun, 55000, Turkey.
FAU - Tuncer, Munir
AU  - Tuncer M
AD  - Department of Molecular Biology and Genetics, Faculty of Science, Ondokuz Mayis 
      University, Samsun, 55000, Turkey.
LA  - eng
GR  - 120Z329/Turkiye Bilimsel ve Teknolojik Arastirma Kurumu/
PT  - Journal Article
DEP - 20231010
PL  - Switzerland
TA  - Mol Biotechnol
JT  - Molecular biotechnology
JID - 9423533
SB  - IM
OTO - NOTNLM
OT  - Characterization
OT  - Cutinolytic esterase
OT  - Expression
OT  - Functional metagenomic
OT  - Plastic degradation
EDAT- 2023/10/10 12:43
MHDA- 2023/10/10 12:43
CRDT- 2023/10/10 11:13
PHST- 2023/08/11 00:00 [received]
PHST- 2023/09/19 00:00 [accepted]
PHST- 2023/10/10 12:43 [medline]
PHST- 2023/10/10 12:43 [pubmed]
PHST- 2023/10/10 11:13 [entrez]
AID - 10.1007/s12033-023-00916-7 [pii]
AID - 10.1007/s12033-023-00916-7 [doi]
PST - aheadofprint
SO  - Mol Biotechnol. 2023 Oct 10. doi: 10.1007/s12033-023-00916-7.