(Below N is a link to NCBI taxonomic web page and E link to ESTHER at designed phylum.) > cellular organisms: NE > Eukaryota: NE > Opisthokonta: NE > Fungi: NE > Dikarya: NE > Ascomycota: NE > saccharomyceta: NE > Pezizomycotina: NE > leotiomyceta: NE > Eurotiomycetes: NE > Eurotiomycetidae: NE > Eurotiales: NE > Trichocomaceae: NE > Thermomyces: NE > Thermomyces lanuginosus: NE
LegendThis sequence has been compared to family alignement (MSA) red => minority aminoacid blue => majority aminoacid color intensity => conservation rate title => sequence position(MSA position)aminoacid rate Catalytic site Catalytic site in the MSA RPVRRAVPQDLLDQFELFSQYSAAAYCAANNHAPVGSDVTCSENVCPEVD AADATFLYSFEDSGLGDVTGLLALDNTNKLIVLSFRGSRSVENWIANLAA DLTEISDICSGCEGHVGFVTSWRSVADTIREQVQNAVNEHPDYRVVFTGH SLGGALATIAAAALRGNGYNIDVFSYGAPRVGNRAFAEFLTAQTGGTLYR ITHTNDIVPRLPPRDWGYSHSSPEYWVTSGNDVPVTANDITVVEGIDSTD GNNQGNIPDIPSHLWYFGPISECD
References
Title: Perspectives on the Role of Enzymatic Biocatalysis for the Degradation of Plastic PET Magalhaes RP, Cunha JM, Sousa SF Ref: Int J Mol Sci, 22:11257, 2021 : PubMed
Plastics are highly durable and widely used materials. Current methodologies of plastic degradation, elimination, and recycling are flawed. In recent years, biodegradation (the usage of microorganisms for material recycling) has grown as a valid alternative to previously used methods. The evolution of bioengineering techniques and the discovery of novel microorganisms and enzymes with degradation ability have been key. One of the most produced plastics is PET, a long chain polymer of terephthalic acid (TPA) and ethylene glycol (EG) repeating monomers. Many enzymes with PET degradation activity have been discovered, characterized, and engineered in the last few years. However, classification and integrated knowledge of these enzymes are not trivial. Therefore, in this work we present a summary of currently known PET degrading enzymes, focusing on their structural and activity characteristics, and summarizing engineering efforts to improve activity. Although several high potential enzymes have been discovered, further efforts to improve activity and thermal stability are necessary.
Due to the rising global environment protection awareness, recycling strategies that comply with the circular economy principles are needed. Polyesters are among the most used materials in the textile industry; therefore, achieving a complete poly(ethylene terephthalate) (PET) hydrolysis in an environmentally friendly way is a current challenge. In this work, a chemo-enzymatic treatment was developed to recover the PET building blocks, namely terephthalic acid (TA) and ethylene glycol. To monitor the monomer and oligomer content in solid samples, a Fourier-transformed Raman method was successfully developed. A shift of the free carboxylic groups (1632 cm(-1) ) of TA into the deprotonated state (1604 and 1398 cm(-1) ) was observed and bands at 1728 and 1398 cm(-1) were used to assess purity of TA after the chemo-enzymatic PET hydrolysis. The chemical treatment, performed under neutral conditions (T = 250 degree C, P = 40 bar), led to conversion of PET into 85% TA and small oligomers. The latter were hydrolysed in a second step using the Humicola insolens cutinase (HiC) yielding 97% pure TA, therefore comparable with the commercial synthesis-grade TA (98%).
Title: Engineering of Thermomyces lanuginosus lipase Lip: creation of novel biocatalyst for efficient biosynthesis of chiral intermediate of Pregabalin Li XJ, Zheng RC, Ma HY, Zheng YG Ref: Applied Microbiology & Biotechnology, 98:2473, 2014 : PubMed
Efficient and highly enantioselective hydrolysis of 2-carboxyethyl-3-cyano-5-methylhexanoic acid ethyl ester (CNDE) is the most crucial step in chemoenzymatic synthesis of Pregabalin. By using site-saturation mutagenesis and high-throughput screening techniques, lipase Lip from Thermomyces lanuginosus DSM 10635 was engineered to improve its activity towards CNDE. The triple mutant, S88T/A99N/V116D exhibited a 60-fold improvement in specific activity for CNDE (2.35 U/mg) over the wild-type Lip (0.039 U/mg). Modeling and docking studies demonstrated that the mutant could more effectively stabilize oxygen anions in transition states and the lid of Lip in the open conformation. Additionally, the kinetic resolution of CNDE catalyzed by Escherichia coli cell overexpressing S88T/A99N/V116D mutant afforded (3S)-2-carboxyethyl-3-cyano-5-methylhexanoic acid in 42.4 % conversion and 98 % ee within 20 h with a substrate loading of 1 M (255 g/l). These results demonstrated that a novel and promising biocatalyst was created for efficient chemoenzymatic manufacturing of Pregabalin.
Thermomyces lanuginosus (Humicola lanuginosa) lipase (lip); Lipozyme TL; LIP_THELA lipolase TLL