Abstract
In this study, the mph gene encoding methyl parathion hydrolase from Pseudomonas sp. WBC-3 was expressed in Yarrowia lipolytica and the expressed methyl parathion hydrolase was displayed on cell surface of Y. lipolytica. The activity of methyl parathion hydrolase displayed on the yeast cells of the transformant Z51 was 59.5 U mg−1 of cell dry cells (450.6 U per mL of the culture) in the presence of 5.0 mM of Co2+. The displayed methyl parathion hydrolase had the optimal pH of 9.5 and the optimal temperature of 40 °C, respectively and was stable in the pH range of 4.5–11 and up to 40 °C. The displayed methyl parathion hydrolase was also stimulated by Co2+, Cu2+, Ni2+ and Mn2+, and was not affected by Fe2+, Fe3+, Na+, K+, Ca2+ and Zn2+, but was inhibited by other cations tested. Under the optimal conditions (OD600nm = 2.6, the substrate concentration = 100 mg L−1 and 40 °C), 90.8 % of methyl parathion was hydrolyzed within 30 min. Under the similar conditions, 98.7, 97.0, 96.5 and 94.4 % of methyl parathion in tap water (pH 9.5), tap water (pH 6.8), seawater (pH 9.5) and natural seawater (pH 8.2) were hydrolyzed, respectively, suggesting that the methyl parathion hydrolase displayed on the yeast cells can effectively remove methyl parathion in water.
Similar content being viewed by others
References
Adams A, Gottschling DE, Kaiser CA, Stearms T (1998) Yeast immunofluorescence. In: Burke D, Dawson D (eds) Methods in yeast genetics: a Cold Spring Harbor Laboratory course manual. Cold Spring Harbor Laboratory, New York, p 100
Chu X, Zhang X, Chen Y, Liu H, Song D (2003) Study on the properties of methyl parathion hydrolase from Pseudomonas sp.W BC-3. Act Microbiol Sin 4:453–459
Efremenko EN, Sergeeva VS (2001) Organophosphate hydrolase- an enzyme catalyzing degradation of phosphorus-containing toxin and pesticides. Russ Chem Bull Intern Ed 50:1826–1832
Fu GP, Cui Z, Huang T, Li SP (2004) Expression, purification, and characterization of a novel methyl parathion hydrolase. Protein Expr Purif 36:170–176
Jolivalt C, Madzak C, Brault A, Caminade E, Malosse C, Mougin C (2005) Expression of laccase lllb from the white-rot fungus Trametes versicolor in the yeast Yarrowia lipolytica for environmental applications. Appl Microbiol Biotechnol 66:450–456
Li CK, Zhu YR, Benz I, Schmidt MA, Chen W, Mulchandani A (2008) Presentation of functional organophosphorus hydrolase fusions on the surface of Escherichia coli by the AIDA-I autotransporter pathway. Biotechnol Bioeng 99:485–490
Liu YH, Liu Y, Chen ZS, Lian J, Huang X, Chung YC (2004) Purification and characterization of a novel organophosphorus pesticide hydrolase from Penicillium lilacinum BP303. Enzyme Microbiol Technol 34:297–303
Liu H, Zhang JJ, Wang SJ, Zhang XE, Zhou NY (2005) Plasmid-borne catabolism of methyl parathion and ρ-nitrophenol in Pseudomonas sp, strain WBC-3. Biochem Biophys Res Commun 334:1107–1114
Liu FY, Hong MZ, Liu DM, Li YW, Shou PS, Yan H, Shi GQ (2007) Biodegradation of methyl parathion by Acinetobacter radioresistens USTB-04. J Environ Sci 19:1257–1260
Madzak C, Gaillardin C, Beckerich JM (2004) Heterologous protein expression and secretion in the non-conventional yeast Yarrowia lipolytica: a review. J Biotechnol 109:63–81
Ni XM, Yue LX, Chi ZM, Li J, Wang XH, Madzak C (2009) Alkaline protease gene cloning from the marine yeast Aureobasidium pullulans HN2-3 and the protease surface display on Yarrowia lipolytica. Mar Biotechnol 11:81–89
Pakala SB, Gorla P, Pinjari AB, Krovidi RK, Baru R, Yanamandra M, Merrick M, Siddavattam D (2007) Biodegradation of methyl parathion and ρ-nitrophenol: evidence for the presence of a ρ-nitrophenol 2-hydroxylase in a Gram-negative Serratia sp. strain DS001. Appl Microbiol Biotechnol 73:1452–1462
Shen YJ, Lu P, Mei H, Yu HJ, Hong Q, Li SP (2009) Isolation of a methyl parathion-degrading strain Stenotrophomonas sp. SMSP-1 and cloning of the ophc2 gene. Biodegradation. doi:10.1007/s10532-010-9343-2
Takayama K, Suye S, Kuroda K, Ueda M, Kitaguchi T, Tsuchiyama K, Fukuda T, Chen W, Mulchandani A (2006) Surface display of organophosphorus hydrolase on Saccharomyces cerevisiae. Biotechnol Prog 22:939–943
Theriot CM, Grunden AM (2011) Hydrolysis of organophosphorus compounds by microbial enzymes. Appl Microbiol Biotechnol 89:35–43
Ueda M, Tanaka A (2000) Genetic immobilization of proteins on the yeast cell surface. Biotechnol Adv 18:121–140
Wu NF, Deng MJ, Liang GY, Chu XY, Yao B, Fan YL (2004) Cloning and expression of ophc2, a new organophosphorus hydrolase gene. Chin Sci Bull 49:1245–1249
Xuan JM, Fournier P, Gaillardin C (1988) Cloning of the LYS5 gene encoding saccharopine dehydrogenase. Curr Genet 14:15–21
Yang W, Zhou YF, Dai HP, Bi LJ, Zhang ZP, Zhang XH, Leng Y, Zhang XE (2008a) Application of methyl parathion hydrolase (MPH) as a labeling enzyme. Anal Bioanal Chem 390:2133–2140
Yang C, Cai N, Dong M, Jiang H, Li JM, Qiao CL (2008b) Surface display of MPH on Pseudomonas putida JS444 using ice nucleation protein and its application in detoxification of organophosphates. Biotechnol Bioeng 99:30–37
Yang C, Zhu Y, Yang J, Liu Z, Qiao CL, Mulchandani A (2008c) Development of an autofluorescent whole-cell biocatalyst by displaying dual functional moieties on Escherichia coli cell surfaces and construction of a coculture with organophosphate-mineralizing activity. Appl Environ Microbiol 74:7733–7739
Yang C, Zhao Q, Liu Z, Li Q, Qiao H, Mulchandania Chen W (2008d) Cell surface display of functional macromolecule fusions on Escherichia coli for development of an autofluorescent whole-cell biocatalyst. Environ Sci Technol 42:6105–6110
Yu H, Yan X, Shen W, Hong Q, Zhang J, Shen Y, Li S (2009) Expression of methyl parathion hydrolase in Pichia pastoris. Curr Microbiol 59:573–578
Yue LX, Chi ZM, Wang L, Liu J, Madzak C, Li J, Wang XH (2008) Construction of a new plasmid for surface display on cells of Yarrowia lipolytica. J Microbiol Method 72:116–123
Zhang ZH, Hong Q, Xu JH, Zhang XZ, Li SP (2006) Isolation of fenitrothion-degrading strain Burkholderia sp. FDS-1 and cloning of mpd gene. Biodegradation 17:275–283
Acknowledgments
This work was supported by Grant 31070029 from National Natural Science Foundation of China, and State Oceanic Administration People’s Republic of China for providing financial support to carry out this work. The Grant No. is 201005032.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Wang, XX., Chi, Z., Ru, SG. et al. Genetic surface-display of methyl parathion hydrolase on Yarrowia lipolytica for removal of methyl parathion in water. Biodegradation 23, 763–774 (2012). https://doi.org/10.1007/s10532-012-9551-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10532-012-9551-z