Pectinases Secretion by Saccharomyces cerevisiae: Optimization in Solid-State Fermentation and Identification by a Shotgun Proteomics Approach

A sequential design strategy was applied to optimize the secretion of pectinases by a Saccharomyces cerevisiae strain, from Brazilian sugarcane liquor vat, on passion fruit residue flour (PFRF), through solid-state fermentation (SSF). A factorial design was performed to determine the influence variables and two rotational central composite designs were executed. The validated experimental result was of 7.1 U mL−1 using 50% PFRF (w/w), pH 5, 30 °C for 24 h, under static SSF. Polygalacturonase, pectin methyl esterase, pectin–lyase and pectate–lyase activities were 3.5; 0.08; 3.1 and 0.8 U mL−1, respectively. Shotgun proteomics analysis of the crude extract enabled the identification of two pectin–lyases, one pectate–lyase and a glucosidase. The crude enzymatic extract maintained at least 80% of its original activity at pH values and temperatures ranging from 2 to 8 and 30 to 80 °C, respectively, over 60 min incubation. Results revealed that PFRF might be a cost-effective and eco-friendly substrate to produce pectinases. Statistical optimization led to fermentation conditions wherein pectin active proteins predominated. To the extent of our knowledge, this is the first study reporting the synthesis of pectate lyase by S. cerevisiae.

[1]  E. Naumova,et al.  Natural Polymorphism of Pectinase PGU Genes in the Saccharomyces Yeasts , 2021, Microbiology.

[2]  M. Eisenacher,et al.  MaCPepDB: A Database to Quickly Access All Tryptic Peptides of the UniProtKB. , 2021, Journal of proteome research.

[3]  Sankha Bhattacharya,et al.  Central Composite Design for Response Surface Methodology and Its Application in Pharmacy , 2021, Response Surface Methodology in Engineering Science [Working Title].

[4]  Ahasanul Karim,et al.  Kluyveromyces marxianus: An emerging yeast cell factory for applications in food and biotechnology. , 2020, International journal of food microbiology.

[5]  E. Costa,et al.  Obtenção e caracterização de vinagre de manga pelo método de acetificação de Orleans , 2020 .

[6]  Juliana John,et al.  Advances in upstream and downstream strategies of pectinase bioprocessing: A review. , 2020, International journal of biological macromolecules.

[7]  N. Melnichuk,et al.  Valorization of two agroindustrial wastes to produce alpha-amylase enzyme from Aspergillus oryzae by solid-state fermentation. , 2020, Waste management.

[8]  S. Samanta MICROBIAL PECTINASES: A REVIEW ON MOLECULAR AND BIOTECHNOLOGICAL PERSPECTIVES , 2019, Journal of Microbiology, Biotechnology and Food Sciences.

[9]  M. T. Neves-Petersen,et al.  Optimized production of Aspergillus aculeatus URM4953 polygalacturonases for pectin hydrolysis in hog plum (Spondias mombin L.) juice , 2019, Process Biochemistry.

[10]  R. Hours,et al.  Production of an endopolygalacturonase from Wickerhanomyces anomalus with disintegration activity on plant tissues , 2019, Biocatalysis and Agricultural Biotechnology.

[11]  Muhammad Bilal,et al.  Recent advances in the production strategies of microbial pectinases-A review. , 2019, International journal of biological macromolecules.

[12]  M. Fan,et al.  Application and optimization of solid-state fermentation process for enhancing polygalacturonase production by Penicillium expansum , 2018 .

[13]  Huimin Zhao,et al.  Recent advances in metabolic engineering of Saccharomyces cerevisiae: New tools and their applications. , 2018, Metabolic engineering.

[14]  F. Bauer,et al.  Agitation impacts fermentation performance as well as carbon and nitrogen metabolism in Saccharomyces cerevisiae under winemaking conditions , 2018 .

[15]  R. Cansian,et al.  Biotechnological potential of agro-industrial waste in the synthesis of pectin lyase from Aspergillus brasiliensis , 2018, Food science and technology international = Ciencia y tecnologia de los alimentos internacional.

[16]  Ashok Pandey,et al.  Production of Pectinase from Bacillus sonorensis MPTD1. , 2018, Food technology and biotechnology.

[17]  C. R. Duarte,et al.  Effects of dehydration methods on quality characteristics of yellow passion fruit co-products. , 2017, Journal of the science of food and agriculture.

[18]  C. R. Duarte,et al.  Impact of freeze‐drying on bioactive compounds of yellow passion fruit residues , 2017 .

[19]  Parameswaran Binod,et al.  Recent advancements in the production and application of microbial pectinases: an overview , 2017, Reviews in Environmental Science and Bio/Technology.

[20]  M. Palma,et al.  Proteome profiling reveals insights into secondary metabolism in Maytenus ilicifolia (Celastraceae) cell cultures producing quinonemethide triterpenes , 2017, Plant Cell, Tissue and Organ Culture (PCTOC).

[21]  R. Simpson,et al.  Effects of Blanching and Hot Air Drying Conditions on the Physicochemical and Technological Properties of Yellow Passion Fruit (Passiflora edulis Var. Flavicarpa) by-Products , 2017 .

[22]  O. Oumer Pectinase: Substrate, Production and their Biotechnological Applications , 2017 .

[23]  Qinyi Zhao On the indirect relationship between protein dynamics and enzyme activity. , 2017, Progress in biophysics and molecular biology.

[24]  A. A. Brasil,et al.  Quantitative proteomic analysis of the Saccharomyces cerevisiae industrial strains CAT-1 and PE-2. , 2017, Journal of proteomics.

[25]  R. Mahajan,et al.  Cost-effective and concurrent production of industrially valuable xylano-pectinolytic enzymes by a bacterial isolate Bacillus pumilus AJK , 2017, Preparative biochemistry & biotechnology.

[26]  X. Lu,et al.  Purification and characterization of exo-polygalacturonase from Zygoascus hellenicus V25 and its potential application in fruit juice clarification , 2016, Food Science and Biotechnology.

[27]  P. Thonart,et al.  Production and Properties of a Thermostable, pH—Stable Exo-Polygalacturonase Using Aureobasidium pullulans Isolated from Saharan Soil of Algeria Grown on Tomato Pomace , 2016, Foods.

[28]  Rajender S. Sangwan,et al.  Bio-processing of Agro-industrial Wastes for Production of Food-grade Enzymes: Progress and Prospects , 2016 .

[29]  N. Krieger,et al.  Production of pectinases by solid-state fermentation of a mixture of citrus waste and sugarcane bagasse in a pilot-scale packed-bed bioreactor , 2016 .

[30]  K. Elst,et al.  Pectin content and composition from different food waste streams. , 2016, Food chemistry.

[31]  S. Gummadi,et al.  Enhanced production of pectinase by Saccharomyces cerevisiae isolate using fruit and agro-industrial wastes: Its application in fruit and fiber processing , 2016 .

[32]  Adrian J Mulholland,et al.  On the Temperature Dependence of Enzyme-Catalyzed Rates. , 2016, Biochemistry.

[33]  J. Kronstad,et al.  Regulation of the fungal secretome , 2016, Current Genetics.

[34]  Dinesh Yadav,et al.  Molecular Biology of Microbial Pectate Lyase: A Review , 2016 .

[35]  Jeremy D O'Connell,et al.  Proteome-wide quantitative multiplexed profiling of protein expression: carbon-source dependency in Saccharomyces cerevisiae , 2015, Molecular biology of the cell.

[36]  J. Andreaus,et al.  The characterization of a pectin-degrading enzyme from Aspergillus oryzae grown on passion fruit peel as the carbon source and the evaluation of its potential for industrial applications , 2015 .

[37]  P. Robinson,et al.  Enzymes: principles and biotechnological applications , 2015, Essays in biochemistry.

[38]  M. Reuss,et al.  Metabolic efficiency in yeast Saccharomyces cerevisiae in relation to temperature dependent growth and biomass yield. , 2015, Journal of thermal biology.

[39]  C. Tarı,et al.  Evaluation of agro-industrial wastes, their state, and mixing ratio for maximum polygalacturonase and biomass production in submerged fermentation , 2015, Environmental technology.

[40]  R. Bhat,et al.  Impact of structural stability of cold adapted Candida antarctica lipase B (CaLB): in relation to pH, chemical and thermal denaturation , 2015 .

[41]  Vijayakumar Poondla,et al.  Low temperature active pectinases production by Saccharomyces cerevisiae isolate and their characterization , 2015 .

[42]  R. Hours,et al.  Production of Pectinolytic Enzymes by the Yeast Wickerhanomyces anomalus Isolated from Citrus Fruits Peels , 2013, Biotechnology research international.

[43]  C. Tarı,et al.  Pectinase enzyme-complex production by Aspergillus spp. in solid-state fermentation: A comparative study , 2012 .

[44]  M. Arévalo-Villena,et al.  Pectinases yeast production using grape skin as carbon source , 2009 .

[45]  S. Gummadi,et al.  Batch and fed batch production of pectin lyase and pectate lyase by novel strain Debaryomyces nepalensis in bioreactor. , 2008, Bioresource technology.

[46]  Glaucia Maria Pastore,et al.  Isolamento e seleção de microrganismos pectinolíticos a partir de resíduos provenientes de agroindústrias para produção de aromas frutais , 2006 .

[47]  Reena Gupta,et al.  Microbial pectinolytic enzymes: A review , 2005 .

[48]  C. J. Robnett,et al.  MOLECULAR RELATIONSHIPS AMONG HYPHAL ASCOMYCETOUS YEASTS AND YEASTLIKE TAXA , 1995 .

[49]  M. Aigle,et al.  Detection of polygalacturonase, pectin‐lyase and pectin‐esterase activities in a Saccharomyces cerevisiae strain , 1994, Yeast.

[50]  D. Pyle,et al.  Endopolygalacturonase production from Kluyveromyces marxianus. I, resolution, purification, and partial characterisation of the enzyme , 1990 .