Abstract
Paraphoma-related fungal strain B47-9 secreted a biodegradable plastic (BP)-degrading enzyme which amounted to 68 % (w/w) of the total secreted proteins in a culture medium containing emulsified poly(butylene succinate-co-adipate) (PBSA) as sole carbon source. The gene for this enzyme was found to be composed of an open reading frame consisting of 681 nucleotides encoding 227 amino acids and two introns. Southern blot analysis showed that this gene exists as a single copy. The deduced amino acid sequence suggested that this enzyme belongs to the cutinase (E.C.3.1.1.74) family; thus, it was named P araphoma-related fungus cutinase-like enzyme (PCLE). It degraded various types of BP films, such as poly(butylene succinate), PBSA, poly(butylene adipate-co-terephthalate), poly(ε-caprolactone), and poly(dl-lactic acid). It has a molecular mass of 19.7 kDa, and an optimum pH and temperature for degradation of emulsified PBSA of 7.2 and 45 °C, respectively. Ca2+ ion at a concentration of about 1.0 mM markedly enhanced the degradation of emulsified PBSA.
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References
Kasuya K, Ishii N, Inoue Y, Yazawa K, Tagaya T, Yotsumoto T, Kazahaya J, Nagai D (2009) Characterization of a mesophilic aliphatic-aromatic copolyester-degrading fungus. Polym Degrad Stab 94(8):1190–1196. doi:10.1016/j.polymdegradstab.2009.04.013
Kitamoto HK, Shinozaki Y, Cao XH, Morita T, Konishi M, Tago K, Kajiwara H, Koitabashi M, Yoshida S, Watanabe T, Sameshima-Yamashita Y, Nakajima-Kambe T, Tsushima S (2011) Phyllosphere yeasts rapidly break down biodegradable plastics. AMB Express 1(1):44. doi:10.1186/2191-0855-1-44
Kodama Y, Masaki K, Kondo H, Suzuki M, Tsuda S, Nagura T, Shimba N, Suzuki E, Iefuji H (2009) Crystal structure and enhanced activity of a cutinase-like enzyme from Cryptococcus sp. strain S-2. Proteins 77(3):710–717. doi:10.1002/prot.22484
Koitabashi M, Noguchi MT, Sameshima-Yamashita Y, Hiradate S, Suzuki K, Yoshida S, Watanabe T, Shinozaki Y, Tsushima S, Kitamoto HK (2012) Degradation of biodegradable plastic mulch films in soil environment by phylloplane fungi isolated from gramineous plants. AMB Express 2:40. doi:10.1186/219-0855-2-40
Koschorreck K, Liu D, Kazenwadel C, Schmid RD, Hauer B (2010) Heterologous expression, characterization and site-directed mutagenesis of cutinase CUTAB1 from Alternaria brassicicola. Appl Microbiol Biotechnol 87:991–997. doi:10.1007/s00253-010-2533-3
Lange K-B (2012) European bioplastics: fivefold growth of the bioplastics market by 2016. Press release, European Bioplastics, Berlin. http://en.european-bioplastics.org/wp-content/uploads/2012/10/PR_market_study_bioplastics_ENG.pdf. Accessed 8 October 2013
Masaki K, Kamini NR, Ikeda H, Iefuji H (2005) Cutinase-like enzyme from the yeast Cryptococcus sp. strain S-2 hydrolyzes polylactic acid and other biodegradable plastics. Appl Environ Microbiol 71(11):7548–7550. doi:10.1128/AEM.71.11.7548-7550.2005
Ohtaki S, Maeda H, Takahashi T, Yamagata Y, Hasegawa F, Gomi K, Nakajima T, Abe K (2006) Novel hydrophobic surface binding protein, HsbA, produced by Aspergillus oryzae. Appl Environ Microbiol 72(4):2407–2413. doi:10.1128/AEm.72.4.2407-2413.2006
Rubio MB, Cardoza RE, Hermosa R, Gutierrez S, Monte E (2008) Cloning and characterization of the Thcut1 gene encoding a cutinase of Trichoderma harzianum T34. Curr Genet 54:301–312. doi:10.1007/s00294-008-0218-6
Sang BI, Hori K, Tanji Y, Unno H (2002) Fungal contribution to in situ biodegradation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) film in soil. Appl Microbiol Biothchnol 58(2):241–247. doi:10.1007/s00253-001-0884-5
Shinozaki Y, Morita T, Cao XH, Yoshida S, Koitabashi M, Watanabe T, Suzuki K, Sameshima-Yamashita Y, Nakajima-Kambe T, Fujii T, Kitamoto HK (2013a) Biodegradable plastic-degrading enzyme from Pseudozyma antarctica: cloning, sequencing, and characterization. Appl Microbiol Biotechnol 97(7):2951–2959. doi:10.1007/s00253-012-4188-8
Shinozaki Y, Watanabe T, Nakajima-Kambe T, Kitamoto HK (2013b) Rapid and simple colorimetric assay for detecting the enzymatic degradation of biodegradable plastic films. J Biosci Bioeng 115(1):111–114. doi:10.1016/j.jbiosc.2012.08.010
Shinozaki Y, Kikkawa Y, Sato S, Fukuoka T, Watanabe T, Yoshida S, Nakajima-Kambe T, Kitamoto HK (2013c) Enzymatic degradation of polyester films by a cutinase-like enzyme from Pseudozyma antarctica: surface plasmon resonance and atomic force microscopy study. Appl Microbiol Biotechnol 97(19):8591–8598. doi:10.1007/s00253-012-4673-0
Sumner C, Sabot A, Turner K, Krause S (2000) A transducer based on enzyme-induced degradation of thin polymer films monitored by surface plasmon resonance. Anal Chem 72(21):5225–5232. doi:10.1021/ac000411y
Suzuki K, Sakamoto H, Shinozaki Y, Tabata J, Watanabe T, Mochizuki A, Koitabashi M, Fujii T, Tsushima S, Kitamoto HK (2013) Affinity purification and characterization of a biodegradable plastic-degrading enzyme from a yeast isolated from the larval midgut of a stag beetle, Aegus laevicollis. Appl Microbiol Biotechnol 97(17):7679–7688. doi:10.1007/s00253-012-4595-x
Takahashi T, Maeda H, Yoneda S, Ohtaki S, Yamagata Y, Hasegawa F, Gomi K, Nakajima T, Abe K (2005) The fungal hydrophobin RolA recruits polyesterase and laterally moves on hydrophobic surface. Mol Microbiol 57(6):1780–1796. doi:10.1111/j.1365-2958.2005.04803.x
Yang S, Xu H, Yan Q, Liu Y, Zhou P, Jiang Z (2013) A low molecular mass cutinase of Thielavia terrestris efficiently hydrolyzes poly(esters). J Ind Microbiol Biotechnol 40:217–226. doi:10.1007/s10295-012-1222-x
Acknowledgments
We thank Showa Denko K. K. for generously supplying the Bionolle materials. We also acknowledge Dr. E. Suto for the valuable comments in this research. This research was financially supported by the Science and Technology Research Promotion Program for Agriculture, Forestry, Fisheries and Food Industry and the National Institute for Agro-Environmental Sciences, Japan.
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Ken Suzuki and Masako Tsujimoto Noguchi contributed equally to this work.
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Suzuki, K., Noguchi, M.T., Shinozaki, Y. et al. Purification, characterization, and cloning of the gene for a biodegradable plastic-degrading enzyme from Paraphoma-related fungal strain B47-9. Appl Microbiol Biotechnol 98, 4457–4465 (2014). https://doi.org/10.1007/s00253-013-5454-0
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DOI: https://doi.org/10.1007/s00253-013-5454-0