Purification-free antifungal biocontrol platform using engineered Saccharomyces cerevisiae secreting Trichoderma atroviride chitinase

Keywords

Saccharomyces cerevisiae / Trichoderma atroviride / Chitinase / Yeast secretion / Antifungal biocontrol / Extracellular enzyme production

References

[1] Abdelraouf AMN, Al-Hazmi NE, Naguib DM, et al.. Streptomyces griseus versus Trichoderma viride chitinase as an anti-inflammatory and antifungal agent against human pathogenic fungi. Indian J Microbiol, 2024, 64: 1863-1869

[2] Adrangi S, Faramarzi MA. From bacteria to human: a journey into the world of chitinases. Biotechnol Adv, 2013, 31: 1786-1795

[3] Ahmad M, Hirz M, Pichler H, Schwab H. Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production. Appl Microbiol Biotechnol, 2014, 98: 5301-5317

[4] Benítez T, Rincón AM, Limón MC, Codón AC. Biocontrol mechanisms of Trichoderma strains. Int Microbiol, 2004, 7: 249-260

[5] Boer H, Teeri TT, Koivula A, et al.. Heterologous expression and site-directed mutagenesis studies of two Trichoderma harzianum chitinases, Chit33 and Chit42, in Escherichia coli. Protein Expr Purif, 2007, 51: 216-226

[6] Božinović M, Sokač Cvetnić T, Šalić A, et al.. Standardization of 3,5-dinitrosalicylic acid (DNS) assay for measuring xylanase activity: detecting and solving problems. Croat J Food Sci Technol, 2023, 15: 151-162

[7] Cabib E, Roh DH, Schmidt M, Crotti LB, Varma A. The yeast cell wall and septum as paradigms of cell growth and morphogenesis. J Biol Chem, 2001, 276: 19679-19682

[8] Carsolio C, Gutiérrez A, Jiménez B, et al.. Characterization of ech42, a Trichoderma harzianum endochitinase gene expressed during mycoparasitism. Proc Natl Acad Sci U S A, 1994, 91: 10903-10907

[9] Cereghino JL, Cregg JM. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev, 2000, 24: 45-66

[10] Csárdi G, Franks A, Choi DS, Airoldi EM, Drummond DA. Accounting for experimental noise reveals that mRNA levels, amplified by post-transcriptional processes, largely determine steady-state protein levels in yeast. PLoS Genet, 2015, 11 Article ID: e1005206

[11] da Silva AF, García Fraga B, López Seijas J, Sieiro C. Optimizing the expression of a heterologous chitinase: a study of different promoters. Bioengineered, 2016, 8: 428-432

[12] Delic M, Rebnegger C, Wanka F, et al.. Oxidative folding and unfolding in recombinant protein production: an underexplored bottleneck in yeast expression systems. Microb Cell Fact, 2013, 12: 89

[13] Deng J-J, Zhang Y, Zhou J, et al.. Heterologous expression and characterization of an antifungal chitinase (Chit46) from Trichoderma harzianum GIM 3.442 and its application in colloidal chitin conversion. Int J Biol Macromol, 2019, 134: 113-121

[14] Draborg H, Christgau S, Halkier T, et al.. Secretion of an enzymatically active Trichoderma harzianum endochitinase by Saccharomyces cerevisiae. Curr Genet, 1996, 29: 404-409

[15] Druzhinina IS, Seidl-Seiboth V, Herrera-Estrella A, et al.. Trichoderma: the genomics of opportunistic success. Nat Rev Microbiol, 2011, 9: 749-759

[16] El Sayed GM, Emam MTH, Hammad MA, Mahmoud SH. Gene cloning, heterologous expression, and in silico analysis of chitinase B from for biocontrol of larvae infesting maize crops. Molecules, 2024, 29 Article ID: 1466

[17] Fisher MC, Hawkins NJ, Sanglard D, Gurr SJ. Threats to global food security from emerging fungal and oomycete crop pathogens. Nat Food, 2020, 1: 332-342

[18] Gancedo JM. Yeast carbon catabolite repression. Microbiol Mol Biol Rev, 1998, 62: 334-361

[19] Gasser B, Mattanovich D. A yeast for all seasons – is Pichia pastoris a suitable chassis organism for future bioproduction?. FEMS Microbiol Lett, 2018, 365 Article ID: fny181

[20] Ge L, Rudolph P. Simultaneous introduction of multiple mutations using overlap extension PCR. Biotechniques, 1997, 22: 28-30

[21] González-Martínez KI, Vázquez-Garcidueñas MS, Herrera-Estrella A, et al.. Polyphasic characterization of the biocontrol potential of a novel strain of Trichoderma atroviride isolated from Central Mexico. J Fungi, 2024, 10: 758

[22] Han KH, Kim JH, Kim WS, Han DM. The snpA, a temperature-sensitive suppressor of npgA1, encodes the eukaryotic translation release factor, eRF1, in Aspergillus nidulans. FEMS Microbiol Lett, 2005, 251: 155-160

[23] Harman GE. Overview of mechanisms and uses of Trichoderma spp. Phytopathology, 2006, 96: 190-194

[24] Harman GE, Howell CR, Viterbo A, Chet I, Lorito M. Trichoderma species—opportunistic, avirulent plant symbionts. Nat Rev Microbiol, 2004, 2: 43-56

[25] Hassan F, Meens J, Jacobsen HJ, Kiesecker H. A family 19 chitinase (Chit30) from Streptomyces olivaceoviridis ATCC 11238 expressed in transgenic pea affects the development of Trichoderma harzianum in vitro. J Biotechnol, 2009, 143: 302-308

[26] Hassan MM, El-Shamy AR, Amin BH, El-Sharkawy RM. Influence of protoplast fusion between two Trichoderma spp. on extracellular enzymes production and antagonistic activity. Biotechnol Biotechnol Equip, 2014, 28: 1014-1023

[27] Heo YJ, Kim DH, Kim TG, Yang MS (2007) Secretory expression of human lysozyme using rice amylase 1A signal peptide in Saccharomyces cerevisiae. J Microbiol Biotechnol 17:847–852.

[28] Idiris A, Tohda H, Kumagai H, Takegawa K. Engineering of protein secretion in yeast: strategies and impact on protein production. Appl Microbiol Biotechnol, 2010, 86: 403-417

[29] Ito H, Fukuda Y, Murata K, Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol, 1983, 153: 163-168

[30] Kim JH, Kim TG, Kim DH, Yang MS. Secretory production of soybean β-conglycinin α′-subunit in Saccharomyces cerevisiae and its antigenicity. J Microbiol Biotechnol, 2008, 18: 1645-1650

[31] Kim J-M, Park S-M, Kim D-H. Heterologous expression of a tannic acid-inducible laccase3 of Cryphonectria parasitica in Saccharomyces cerevisiae. BMC Biotechnol, 2010, 10 Article ID: 18

[32] Kim J-M, Song H-Y, Choi H-J, et al.. Characterization of NpgA, a 4′-phosphopantetheinyl transferase of Aspergillus nidulans, and evidence of its involvement in fungal growth and formation of conidia and cleistothecia for development. J Microbiol, 2015, 53: 21-31

[33] Kubicek CP, Steindorff AS, Chenthamara K, et al.. Evolution and comparative genomics of the most common Trichoderma species. BMC Genomics, 2019, 20 Article ID: 485

[34] Kuranda MJ, Robbins PW. Chitinase is required for cell separation during growth of Saccharomyces cerevisiae. J Biol Chem, 1991, 266: 19758-19767

[35] Lee SY, Kim TG, Yang MS (2006) Secretory expression of a horseradish peroxidase by using the rice amylase 1A signal peptide in Saccharomyces cerevisiae. J Microbiol 44:87–91.

[36] Li S, Zhang B, Zhu H-M, Zhu T-H. Cloning and expression of the chitinase encoded by ChiKJ406136 from Streptomyces sampsonii (Millard & Burr) Waksman KJ40 and its antifungal effect. Forests, 2018, 9: 699

[37] Liu Z, Tyo KEJ, Martínez JL, Petranovic D, Nielsen J. Different expression systems for production of recombinant proteins in Saccharomyces cerevisiae. Biotechnol Bioeng, 2016, 113: 1232-1243

[38] Márquez-Fernández O, Trigos A, Ramos-Balderas JL, et al.. Phosphopantetheinyl transferase CfwA/NpgA is required for Aspergillus nidulans secondary metabolism and asexual development. Eukaryot Cell, 2007, 6: 710-720

[39] Nguyen N-L, Kim Y-H, Kim D-H, et al.. Expression and characterization of an M cell-specific ligand-fused dengue virus tetravalent epitope using Saccharomyces cerevisiae. J Biosci Bioeng, 2015, 119: 19-27

[40] Pérez-Martínez AS, Gutiérrez-Sánchez G, Ruiz-Herrera J, et al.. Overexpression, purification and characterization of the Trichoderma atroviride endochitinase, Ech42, in Pichia pastoris. Protein Expr Purif, 2007, 55: 183-188

[41] Pérez-Martínez AS, Ruiz-Herrera J, Martínez-Trujillo A. Heterologous expression of Trichoderma atroviride endochitinase ech42 in Pichia pastoris at low and high dissolved oxygen tensions. Rev Mex Ing Quim, 2014, 13: 93-101

[42] Pontecorvo G, Roper JA, Hemmons LM, Macdonald KD, Bufton AWJ. The genetics of Aspergillus nidulans. Adv Genet, 1953, 5: 141-238

[43] Rosano GL, Ceccarelli EA (2014) Recombinant protein expression in Escherichiacoli: advances and challen ges. Front Microbiol 5:172. https://doi.org/10.3389/fmicb.2014.00172

[44] Shin YJ, Lee SY, Kim TG, Yang MS. Secretory expression of horseradish peroxidase and tobacco anionic peroxidase in Saccharomyces cerevisiae. Protein Expr Purif, 2004, 38: 146-151

[45] Shin M-K, Kim TG, Yang MS. Induction of protective immune responses against challenge of Actinobacillus pleuropneumoniae by oral administration with Saccharomyces cerevisiae expressing Apx toxins in pigs. Vet Immunol Immunopathol, 2013, 151: 132-139

[46] Shukla AA, Thömmes J. Recent advances in large-scale production of monoclonal antibodies and related proteins. Trends Biotechnol, 2010, 28: 253-261

[47] So K-K, Kim D-H, Kang J-Y, et al.. Heterologous expression of Laccase1 from Cryphonectria parasitica in Saccharomyces cerevisiae. Mycobiology, 2025, 53: 36-46

[48] Stubbs AC, Martin KS, Coeshott C et al (2001) Whole recombinant yeast vaccine activates dendritic cells and elicits protective cell-mediated immunity. Nat Med 7:625–629. https://doi.org/10.1038/87974

[49] Subramanian K, Thakur V, Kumar R, Singh BP. Bioconversion of chitin and concomitant production of chitinase and N-acetylglucosamine by novel Achromobacter xylosoxidans isolated from shrimp waste disposal area. Sci Rep, 2020, 10 Article ID: 11898

[50] Swiontek Brzezinska M, Jankiewicz U, Burkowska A, Walczak M. Chitinolytic microorganisms and their possible application in environmental protection. Curr Microbiol, 2014, 68: 71-81

[51] Viterbo A, Haran S, Friesem D, Ramot O, Chet I. Antifungal activity of a novel endochitinase gene (chit36) from Trichoderma harzianum Rifai TM. FEMS Microbiol Lett, 2001, 200: 169-174

[52] Wang S, Li H, Liu S, et al.. High-efficiency secretion and directed evolution of chitinase BcChiA1 in Bacillus subtilis for the conversion of chitinaceous wastes into chitooligosaccharides. Front Bioeng Biotechnol, 2020, 8 Article ID: 432

[53] Wang W, Zhang Y, Li H, Liu S, Qi G. Chitin substrate–mediated stabilization of extracellular chitinases and enhancement of antifungal activity. Int J Biol Macromol, 2020, 164: 3960-3969

[54] Wu J-D, Chou J-C. Optimization of protoplast preparation and regeneration of medical fungus Antrodia cinnamomea. Mycobiology, 2019, 47: 483-493

[55] Yu P, Yan Y, Gu Q, Wang X. Codon optimisation improves the expression of Trichoderma viride sp. endochitinase in Pichia pastoris. Sci Rep, 2013, 3 Article ID: 3043

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