A Novel Endo-Polygalacturonase from Penicillium oxalicum: Gene Cloning, Heterologous Expression and Its Use in Acidic Fruit Juice Extraction
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An endo-polygalacturonase (endo-PGase) exhibiting excellent performance during acidic fruit juice production would be highly attractive to the fruit juice industry. However, candidate endo-PGases for this purpose have rarely been reported. In this study, we expressed a gene from Penicillium oxalicum in Pichia pastoris. The recombinant enzyme PoxaEnPG28C had an optimal enzyme activity at pH 4.5 and 45°C and was stable at pH 3.0–6.5 and < 45°C. The enzyme had a specific activity of 4,377.65 ± 55.37 U/mg towards polygalacturonic acid, and the Km and Vmax values of PoxaEnPG28C were calculated as 1.64 g/l and 6127.45 U/mg, respectively. PoxaEnPG28C increased the light transmittance of orange, lemon, strawberry and hawthorn juice by 13.9 ± 0.3%, 29.4 ± 3.8%, 95.7 ± 10.2% and 79.8 ± 1.7%, respectively; it reduced the viscosity of the same juices by 25.7 ± 1.6%, 52.0 ± 4.5%, 48.2 ± 0.7% and 80.5 ± 2.3%, respectively, and it increased the yield of the juices by 24.5 ± 0.7%, 12.7 ± 2.2%, 48.5 ± 4.2% and 104.5 ± 6.4%, respectively. Thus, PoxaEnPG28C could be considered an excellent candidate enzyme for acidic fruit juice production. Remarkably, fruit juice production using hawthorn as an material was reported for the first time.
Abstract
An endo-polygalacturonase (endo-PGase) exhibiting excellent performance during acidic fruit juice production would be highly attractive to the fruit juice industry. However, candidate endo-PGases for this purpose have rarely been reported. In this study, we expressed a gene from Penicillium oxalicum in Pichia pastoris. The recombinant enzyme PoxaEnPG28C had an optimal enzyme activity at pH 4.5 and 45°C and was stable at pH 3.0–6.5 and < 45°C. The enzyme had a specific activity of 4,377.65 ± 55.37 U/mg towards polygalacturonic acid, and the Km and Vmax values of PoxaEnPG28C were calculated as 1.64 g/l and 6127.45 U/mg, respectively. PoxaEnPG28C increased the light transmittance of orange, lemon, strawberry and hawthorn juice by 13.9 ± 0.3%, 29.4 ± 3.8%, 95.7 ± 10.2% and 79.8 ± 1.7%, respectively; it reduced the viscosity of the same juices by 25.7 ± 1.6%, 52.0 ± 4.5%, 48.2 ± 0.7% and 80.5 ± 2.3%, respectively, and it increased the yield of the juices by 24.5 ± 0.7%, 12.7 ± 2.2%, 48.5 ± 4.2% and 104.5 ± 6.4%, respectively. Thus, PoxaEnPG28C could be considered an excellent candidate enzyme for acidic fruit juice production. Remarkably, fruit juice production using hawthorn as an material was reported for the first time.
참고문헌 (47)
- Willats WGT, "Pectin : new insights into an old polymer are starting to gel", Trends Food Sci. Technol, 17, pp.97~104 (2006)
- Voragen AGJ, "Pectin, a versatile polysaccharide present in plant cell walls", Struct. Chem, 20, pp.263~275 (2009)
- Jayani RS, "Microbial pectinolytic enzymes : a review", Process Biochem, 40, pp.2931~2944 (2005)
- John J, "Advances in upstream and downstream strategies of pectinase bioprocessing : a review", Int. J. Biol. Macromol, 162, pp.1086~1099 (2020)
- Xian L, "Identification and characterization of an acidic and acid-stable endoxyloglucanase from Penicillium oxalicum", Int. J. Biol. Macromol, 86, pp.512~518 (2016)
- Kashyap DR, "Applications of pectinases in the commercial sector : a review", Bioresour. Technol, 77, pp.215~227 (2001)
- Lozano JE, "Fruit manufacturing: scientific basis, engineering properties, and deteriorative reactions of technological importance (Food Engineering Series)" (2006), Springer
- Saúco VG, "Tropical and Subtropical Fruits. Horticulture: Plants for people and places, Volume 1" (2014)
- Cheng Z, "Development of an innovative process for high-temperature fruit juice extraction using a novel thermophilic endo-polygalacturonase from Penicillium oxalicum", Front. Microbiol, 11, pp.1200~ (2020)
- Liu GD, "Genomic and secretomic analyses reveal unique features of the lignocellulolytic enzyme system of Penicillium decumbens", PLoS One, 8, pp.55185~ (2013)
- Zhong Cheng, "A Novel Acid-Stable Endo-Polygalacturonase from Penicillium oxalicum CZ1028: Purification, Characterization, and Application in the Beverage Industry", Journal of Microbiology and Biotechnology, 26(6), pp.989~998 (2016), 한국미생물·생명공학회[DOI]
- Cheng Z, "Identification of an acidic endo-polygalacturonase from Penicillium oxalicum CZ1028 and its broad use in major tropical and subtropical fruit juices production", J. Biosci. Bioeng, 123, pp.665~672 (2017)
- U. K. LAEMMLI, "Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4", Nature, 227(5259), pp.680~685 (1970), Springer Science and Business Media LLC[DOI]
- Bradford MM, "A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding", Anal. Biochem, 72, pp.248~254 (1976)
- Ishida Y, "Cloning and characterization of a polygalacturonase-encoding gene from Penicillium janthinellum", J. Ferment. Bioeng, 84, pp.257~260 (1997)
- van Santen Y, "1.68-A crystal structure of endopolygalacturonase II from Aspergillus niger and identification of active site residues by site-directed mutagenesis", J. Biol. Chem, 274, pp.30474~30480 (1999)
- Liu MQ, "Cloning, expression of Aspergillus niger JL-15 endo-polygalacturonase A gene in Pichia pastoris and oligo-galacturonates production", Protein Expr. Purif, 94, pp.53~59 (2014)
- Huoxiang Zhou, "Secretory Expression and Characterization of an Acidic Endo-Polygalacturonase from Aspergillus niger SC323 in Saccharomyces cerevisiae", Journal of Microbiology and Biotechnology, 25(7), pp.999~1006 (2015), 한국미생물·생명공학회[DOI]
- Yuan P, "A novel acidic and low-temperature-active endo-polygalacturonase from Penicillium sp. CGMCC 1669 with potential for application in apple juice clarification", Food Chem, 129, pp.1369~1375 (2011)
- Yang J, "Cloning, expression and characterization of an acidic endopolygalacturonase from Bispora sp. MEY-1 and its potential application in juice clarification", Process Biochem, 46, pp.272~277 (2011)
- Rojas NL, "Production of heterologous polygalacturonase I from Aspergillus kawachii in Saccharomyces cerevisiae in batch and fed-batch cultures", J. Ind. Microbiol. Biotechnol, 38, pp.1437~1447 (2011)
- Tu T, "Molecular characterization of a thermophilic endo-polygalacturonase from Thielavia arenaria XZ7 with high catalytic efficiency and application potential in the food and feed industries", J. Agr. Food Chem, 62, pp.12686~12694 (2014)
- Sassi AH, "A low-temperature polygalacturonase from P. occitanis: characterization and application in juice clarification", Int. J. Biol. Macromol, 91, pp.158~164 (2016)
- Li YQ, "Two acidic, thermophilic GH28 polygalacturonases from Talaromyces leycettanus JCM 12802 with application potentials for grape juice clarification", Food Chem, 237, pp.997~1003 (2017)
- Tan HD, "Identification and characterization of thermostable endopolygalacturonase II B from Aspergillus luchuensis", J. Food Biochem, 44, pp.e13133~ (2020)
- Tu T, "High-yield production of a low-temperature-active polygalacturonase for papaya juice clarification", Food Chem, 141, pp.2974~2981 (2013)
- Wang JJ, "Efficient expression of an acidic endo-polygalacturonase from Aspergillus niger and its application in juice production", J. Agr. Food Chem, 65, pp.2730~2736 (2017)
- Kant S, "Purification and physicochemical properties of polygalacturonase from Aspergillus niger MTCC 3323", Protein Exp. Purif, 87, pp.11~16 (2013)
- Juwon AD, "Purification, characterization and application of polygalacturonase from Aspergillus niger CSTRF", Malaysian J. Microbiol, 8, pp.175~183 (2012)
- Dinu D, "Enzymes with new biochemical properties in the pectinolytic complex produced by Aspergillus niger MIUG 16", J. Biotechnol, 131, pp.128~137 (2007)
- Guo CT, "Purification and partial characterization of an endo-polygalacturonase from Aspergillus niger", J. Food Biochem, 26, pp.253~265 (2002)
- Singh SA, "A simple fractionation protocol for, and a comprehensive study of the molecular properties of, two major endopolygalacturonases from Aspergillus niger", Biotechnol. Appl. Biochem, 35, pp.115~123 (2002)
- Munir M, "Purification, characterization, kinetics and thermodynamic analysis of polygalacturonase from Aspergillus tamarii for industrial applications", Rev. Mex. Ing. Quím, 19, pp.293~304 (2020)
- Massa C, "Isolation, heterologous expression and characterization of an endopolygalacturonase produced by the phytopathogen Burkholderia cepacia", Protein Exp. Purif, 54, pp.300~308 (2007)
- Tounsi H, "Catalytic properties of a highly thermoactive polygalacturonase from the mesophilic fungus Penicillium occitanis and use in juice clarification", J. Mol. Catal. B-Enzym, 127, pp.56~66 (2016)
- Pan X, "Biochemical characterization of three distinct polygalacturonases from Neosartorya fischeri P1", Food Chem, 188, pp.569~575 (2015)
- Muttalip Gundogdu, "Organic acids, sugars, vitamin C content and some pomological characteristics of eleven hawthorn species (Crataegus spp.) from Turkey", Biological Research, 47(1) (2014), Springer Science and Business Media LLC[DOI]
- Byaruagaba-Bazirake GW, "Characteristics of enzyme-treated banana juice from three cultivars of tropical and subtropical Africa", Afr. J. Food Sci. Technol, 3, pp.277~290 (2012)
- Ali ZM, "A comparative study on wall degrading enzymes, pectin modifications and softening during ripening of selected tropical fruits", Plant Sci, 167, pp.317~327 (2004)
- Xu H, "Overexpression and biochemical characterization of an Endo-α-1, 4-polygalacturonase from Aspergillus nidulans in Pichia pastoris", Int. J. Mol. Sci, 21, pp.2100~ (2020)
- Si-Yuan Zhu, "Improved Homologous Expression of the Acidic Lipase from Aspergillus niger", Journal of Microbiology and Biotechnology, 30(2), pp.196~205 (2020), 한국미생물·생명공학회
- Sanjay Kumar, "Effect of Substrate and Fermentation Conditions on Pectinase and Cellulase Production by Aspergillus niger NCIM 548 in Submerged (SmF) and Solid State Fermentation (SSF)", Food Science and Biotechnology, 20(5), pp.1289~1298 (2011), 한국식품과학회
- 정은비, "Biotransformation of Protopanaxadiol- Type Ginsenosides in Korean Ginseng Extract into Food-Available Compound K by an Extracellular Enzyme from Aspergillus niger", Journal of Microbiology and Biotechnology, 30(10), pp.1560~1567 (2020), 한국미생물·생명공학회[DOI]
- Sanath Kondaveeti, "Bioelectrochemical Detoxification of Phenolic Compounds during Enzymatic Pre-Treatment of Rice Straw", Journal of Microbiology and Biotechnology, 29(11), pp.1760~1768 (2019), 한국미생물·생명공학회[DOI]
- Barman S, "Pectinase production by Aspergillus niger using banana(Musa balbisiana)peel as substrate and its effect on clarification of banana juice", J. Food Sci. Technol, 52, pp.3579~3589 (2015)
- Yang C, "Low-cost recycling production of pectinase to increase the yield and quality of Muzao jujube juice by Aspergillus niger", Biofuel. Bioprod. Bioref, 14, pp.104~116 (2020)
- Pel HJ, "Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88", Nat. Biotechnol, 25, pp.221~231 (2007)
