Characterization of a poly(butylene adipate-co-terephthalate)-  hydrolyzing lipase from Pelosinus fermentans.
Certain α/β hydrolases have the ability to hydrolyze synthetic polyesters. While their partial hydrolysis has a potential for surface functionalization, complete hydrolysis allows recycling of valuable building blocks. Although knowledge about biodegradation of these materials is important regarding their fate in the environment, it is currently limited to aerobic organisms. A lipase from the anaerobic groundwater organism Pelosinus fermentans DSM 17108(PfL1) was cloned and expressed in Escherichia coli BL21-Gold (DE3) and purified from the cell extract. Biochemical characterization with small substrates showed thermoalkalophilic properties (Topt=50 °C, pHopt=7.5) and higher activity towards para-nitrophenyl octanoate (12.7 U mg(-1)) compared to longer and shorter chain lengths (C14 0.7 U mg(-1) and C2 4.3 U mg(-1), respectively). Crystallization and determination of the 3-D structure displayed the presence of a lid structure and a zinc ion surrounded by an extra domain. These properties classify the enzyme into the I.5 lipase family. PfL1 is able to hydrolyze poly(1,4-butylene adipate-co-terephthalate) (PBAT) polymeric substrates. The hydrolysis of PBAT showed the release of small building blocks as detected by liquid chromatography mass spectrometry (LC-MS). Protein dynamics seem to be involved with lid opening for the hydrolysis of PBAT by PfL1.
Institute of Environmental Biotechnology, University of Natural Resources and Life Science (BOKU), Konrad Lorenz Strasse 22, 3430, Tulln an der Donau, Austria.
Antonino
A
Biundo
Austrian Centre for Industrial Biotechnology (ACIB), Petersgasse 14, 8010, Graz, Austria.|Institute of Molecular Bioscience, University of Graz, Universitaetsstraße 15, 8010, Graz, Austria.
Altijana
A
Hromic
Austrian Centre for Industrial Biotechnology (ACIB), Petersgasse 14, 8010, Graz, Austria.|Institute of Molecular Bioscience, University of Graz, Universitaetsstraße 15, 8010, Graz, Austria.
Tea
T
Pavkov-Keller
Austrian Centre for Industrial Biotechnology (ACIB), Petersgasse 14, 8010, Graz, Austria.|Institute of Molecular Bioscience, University of Graz, Universitaetsstraße 15, 8010, Graz, Austria.
Karl
K
Gruber
Institute of Environmental Biotechnology, University of Natural Resources and Life Science (BOKU), Konrad Lorenz Strasse 22, 3430, Tulln an der Donau, Austria.
Felice
F
Quartinello
Austrian Centre for Industrial Biotechnology (ACIB), Konrad Lorenz Strasse 22, 3430, Tulln an der Donau, Austria.
Karolina
K
Haernvall
Austrian Centre for Industrial Biotechnology (ACIB), Konrad Lorenz Strasse 22, 3430, Tulln an der Donau, Austria.
Veronika
V
Perz
Institute of Life Technologies, University of Applied Sciences Western Switzerland (HES-SO Valais/Wallis), Rue du Rawil 47, 1950, Sion, Switzerland.
Miriam S
MS
Arrell
Institute of Life Technologies, University of Applied Sciences Western Switzerland (HES-SO Valais/Wallis), Rue du Rawil 47, 1950, Sion, Switzerland.
Manfred
M
Zinn
Institute of Environmental Biotechnology, University of Natural Resources and Life Science (BOKU), Konrad Lorenz Strasse 22, 3430, Tulln an der Donau, Austria. doris.ribitsch@acib.at.|Austrian Centre for Industrial Biotechnology (ACIB), Konrad Lorenz Strasse 22, 3430, Tulln an der Donau, Austria. doris.ribitsch@acib.at.
Doris
D
Ribitsch
Institute of Environmental Biotechnology, University of Natural Resources and Life Science (BOKU), Konrad Lorenz Strasse 22, 3430, Tulln an der Donau, Austria.|Austrian Centre for Industrial Biotechnology (ACIB), Konrad Lorenz Strasse 22, 3430, Tulln an der Donau, Austria.
Georg M
GM
Guebitz
D000595:Amino Acid Sequence; D000693:Anaerobiosis; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D000068536:Bacillota / Q000201:enzymology* / Q000302:isolation & purification*; D015870:Gene Expression; D060587:Groundwater / Q000382:microbiology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D011091:Polyesters / Q000378:metabolism*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013696:Temperature
D004269:DNA, Bacterial; D011091:Polyesters; D011994:Recombinant Proteins; C488797:poly(butylene adipate-co-butylene terephthalate); D008049:Lipase