In vitro evaluation of probiotic potential and enzymatic profiling of Pichia kudriavzevii Y33 isolated from traditional home-made mango pickle

Prem Lata1, Reena Kumari1, Kiran Bala Sharma1, Shailja Rangra1, Savitri1
1Department of Biotechnology, Himachal Pradesh University, Shimla, India

Tóm tắt

Fermented foods are the results of metabolic activities of various microorganisms. People have traditionally known how to culture desirable microorganisms, primarily lactic acid bacteria, yeasts, and filamentous molds, for the manufacture of edible foods. Yeast isolated from home-made mango pickle from Hamirpur, Himachal Pradesh, was assessed for probiotic properties and their enzymatic profiling. Four yeast isolates were isolated out of which P. kudriavzevii Y33 was selected on the basis of high acid tolerance as well as broadest antimicrobial activity. The selected isolate was observed to have high acid tolerance at pH 2 and show strong antimicrobial activity against all the pathogens examined. P. kudriavzevii Y33 can also withstand high bile concentration and showed high viability index, i.e., 95% at concentration of 2% of bile. The isolate was able to demonstrate high cholesterol assimilation in medium containing ox bile and taurocholate, at 88.58 and 86.83%, respectively. The autoaggregation ability of isolate increases with increasing the time of incubation and showed 87% of autoaggregation after 24 h of incubation. P. kudriavzevii Y33 exhibited resistance towards different antibiotics, found to be positive for exopolysaccharide production and showed no hemolytic activity. The isolate was observed to produce several enzymes such as β-galactosidase, protease, amylase, phytase, and lipase. The results of the current study revealed that P. kudriavzevii Y33 has various beneficial qualities that suggest it could be used as probiotics. Enzymes produced by yeast isolate help in improving flavor and mineral availability in the fermented products.

Tài liệu tham khảo

Alakeji TP, Oloke JK (2020) Association of probiotic potential of strains of Pichia kudriavzevii isolated from Ogi with the number of open reading frame (ORF) in the nucleotide sequences. Afr J Biotecnol 19:148–155. https://doi.org/10.5897/AJB2019.16814 Bae HD, Yanke LJ, Cheng KJ, Selinger LB (1999) A novel staining method for detecting phytase activity. J Microbiol Methods 39:17–22. https://doi.org/10.1016/S0167-7012(99)00096-2 Bajaj B, Raina S, Signh S (2013) Killer toxin from a novel killer yeast Pichia kudriavzevii RY55 with idiosyncratic antibacterial activity. J Basic Microbiol 53:645–656. https://doi.org/10.1002/jobm.201200187 Bajwa J, Sharma N (2018) Evaluation of probiotic properties of yeasts isolated from Sidra–an ethnic fermented fish product of north East India. Int J Curr Microbiol App Sci 7:2632–2643. https://doi.org/10.20546/ijcmas.2018.702.320 Buzzini P, Martini A (2002) Extracellular enzymatic activity profiles in yeast and yeast-like strains isolated from tropical environments. J Appl Microbiol 93:1020–1025. https://doi.org/10.1046/j.1365-2672.2002.01783.x Chakraborty R, Roy S (2018) Exploration of the diversity and associated health benefits of traditional pickles from the Himalayan and adjacent hilly regions of Indian subcontinent. J Food Sci Technol 55:1599–1613. https://doi.org/10.1007/s13197-018-3080-7 Chan GF, Gan HM, Ling HL, Rashid NA (2012) Genome sequence of Pichia kudriavzevii M12, a potential producer of bioethanol and phytase. Eukaryot Cell 11:10. https://doi.org/10.1128/EC.00229-12 Ciszek-Lenda M, Nowak B, Srottek M, Gamian A, Marcinkiewicz J (2011) Immunoregulatory potential of exopolysaccharide from lactobacillus rhamnosus KL37. Effects on the production of inflammatory mediators by mouse macrophages. Int J Exp Pathol 92:382–391. https://doi.org/10.1002/star.201000099 Collado MC, Meriluoto J, Salminen S (2008) Adhesion and aggregation properties of probiotic and pathogen strains. Eur Food Res Technol 226:1065–1073. https://doi.org/10.1007/s00217-007-0632-x Del Re B, Sgorbati B, Miglioli M, Palenzona D (2000) Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum. Lett Appl Microbiol 31:438–442. https://doi.org/10.1046/j.1365-2672.2000.00845.x Delgado-Ospina J, Triboletti S, Alessandria V, Serio A, Sergi M, Paparella A, Rantsiou K, Chaves-López C (2020) Functional biodiversity of yeasts isolated from Colombian fermented and dry cocoa beans. Microorganisms 8:1–17. https://doi.org/10.3390/microorganisms8071086 Filho-Lima JV, Vieira EC, Nicoli JR (2000) Antagonistic effect of lactobacillus acidophilus. Saccharomyces boulardii and Escherichia coli combinations against experimental infections with Shigella flexneri and salmonella enteritidis subsp. typhimurium in gnotobiotic mice. J Appl Microbiol 88:365–370. https://doi.org/10.1046/j.1365-2672.2000.00973.x Furukawa S, Watanabe T, Toyama H, Morinaga Y (2013) Significance of microbial symbiotic coexistence in traditional fermentation. J Biosci Bioeng 116:533–539. https://doi.org/10.1016/j.jbiosc.2013.05.017 Gedek BR (1999) Adherence of Escherichia coli sero group O 157 and the salmonella typhimurium mutant DT 104 to the surface of saccharomyces boulardii. Mycoses 42:261–264. https://doi.org/10.1046/j.1439-0507.1999.00449.x Gilliland SE, Staley TE, Bush LJ (1984) Importance of bile tolerance of lactobacillus acidophilus used as a dietary adjunct. J Dairy Sci 67:3045–3051. https://doi.org/10.3168/jds.S0022-0302(84)81670-7 Helmy EA, Soliman SA, Abdel-Ghany TM, Ganash M (2019) Evaluation of potentially probiotic attributes of certain dairy yeast isolated from buffalo sweetened Karish cheese. Heliyon 5:1–9. https://doi.org/10.1016/j.heliyon.2019.e01649 Hickson M (2011) Probiotics in the prevention of antibiotic-associated diarrhoea and Clostridium difficile infection. Thera Adv Gastroenterol 4:185–197. https://doi.org/10.1177/1756283X11399115 Hill C, Guarner F, Reid G, Gibson GR, Merenstein DJ, Pot B, Morelli L, Canani RB, Flint HJ, Salminen S, Calder PC (2014) The international scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastroenterol Hepatol 11:506–514. https://doi.org/10.1038/nrgastro.2014.66 Kanmani P, Suganya K, Yuvaraj N, Pattukumar V, Paari KA, Arul V (2013) Synthesis and functional characterization of antibiofilm exopolysaccharide produced by enterococcus faecium MC13 isolated from the gut of fish. Appl Biochem Biotechnol 169:1001–1015. https://doi.org/10.1007/s12010-012-0074-1 Kathade S, Aswani M, Nirichan B (2020) Probiotic characterization and cholesterol assimilation ability of Pichia kudriavzevii isolated from the gut of the edible freshwater snail “Pila globosa”. Egypt J Aquat Biol Fish 24:23–39. https://doi.org/10.21608/ejabf.2020.119039 Kordel M, Hofmann B, Schomburg D, Schmid RD (1991) Extracellular lipase of pseudomonas sp. strain ATCC 21808: purification, characterization, crystallization, and preliminary X-ray diffraction data. J Bacteriol 173:4836–4841. https://doi.org/10.1128/jb.173.15.4836-4841.1991 Kwofie MK, Bukari N, Adeboye O (2020) Probiotics potential of yeast and lactic acid bacteria fermented foods and the impact of processing: a review of indigenous and continental food products. Adv Appl Microbiol 10:492–507. https://doi.org/10.4236/aim.2020.109037 Li M, Wang Y, Cui H, Li Y, Sun Y, Qiu H (2020) Characterization of lactic acid bacteria isolated from the gastrointestinal tract of a wild boar as potential probiotics. Front Vet Sci 7:49. https://doi.org/10.3389/fvets.2020.00049 Liong MT, Shah NP (2005) Optimization of cholesterol removal by probiotics in the presence of prebiotics by using a response surface method. Appl Environ Microbiol 71:1745–1753. https://doi.org/10.1128/AEM.71.4.1745-1753.2005 Lombardi A, Dal Maistro L, De Dea P, Gatti M, Giraffa G, Neviani E (2002) A polyphasic approach to highlight genotypic and phenotypic diversities of lactobacillus helveticus strains isolated from dairy starter cultures and cheeses. J Dairy Res 69:139–149. https://doi.org/10.1017/s0022029901005349 Manachini PL, Fortina MG, Parini C (1988) Thermostable alkaline protease produced by bacillus thermoruber—a new species of bacillus. Appl Microbiol Biotechnol 28:409–413. https://doi.org/10.1007/BF00268205 Menezes AG, Ramos CL, Cenzi G, Melo DS, Dias DR, Schwan RF (2020) Probiotic potential, antioxidant activity, and phytase production of indigenous yeasts isolated from indigenous fermented foods. Probiotics Antimicrob Proteins 12:280–288. https://doi.org/10.1007/s12602-019-9518-z Merchán AV, Benito MJ, Galván AI, de Herrera SR (2020) Identification and selection of yeast with functional properties for future application in soft paste cheese. LWT 124:109173. https://doi.org/10.1016/j.lwt.2020.109173 Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428. https://doi.org/10.1021/ac60147a030 Miller JH (1972) Assay of β-galactosidase. In: Experiments in molecular genetics. Cold Spring Harbour Laboratory Press, New York Mishra V, Prasad DN (2005) Application of in vitro methods for selection of lactobacillus casei strains as potential probiotics. Int J Food Microbiol 103:109–115. https://doi.org/10.1016/j.ijfoodmicro.2004.10.047 Mogrovejo DC, Perini L, Gostinčar C, Sepčić K, Turk M, Ambrožič-Avguštin J, Brill FHH, Gunde-Cimerman N (2020) Prevalence of antimicrobial resistance and hemolytic phenotypes in culturable arctic bacteria. Front Microbiol 11:1–13. https://doi.org/10.3389/fmicb.2020.00570 Moon SH, Chang M, Kim HY, Chang HC (2014) Pichia kudriavzevii is the major yeast involved in film-formation, off-odor production, and texture-softening in over-ripened Kimchi. Food Sci Biotechnol 23:489–497. https://doi.org/10.1007/s10068-014-0067-7 Mora D, Fortina MG, Parini C, Ricci G, Gatti M, Giraffa G, Manachini PL (2002) Genetic diversity and technological properties of Streptococcus thermophilus strains isolated from dairy products. J Appl Microbiol 93:278–287. https://doi.org/10.1046/j.1365-2672.2002.01696.x Moser SA, Savage DC (2001) Bile salt hydrolase activity and resistance to toxicity of conjugated bile salts are unrelated properties in lactobacilli. Appl Environ Microbiol 67:3476–3480. https://doi.org/10.1128/AEM.67.8.3476-3480.2001 Ouwehand AC, Salminen S, Isolauri E (2002) Probiotics: an overview of beneficial effects. Antonie Leeuwenhoek 82:279–289. https://doi.org/10.1023/A:1020620607611 Pinto A, Barbosa J, Albano H, Isidro J, Teixeira P (2020) Screening of bacteriocinogenic lactic acid bacteria and their characterization as potential probiotics. Microorganisms 8:393. https://doi.org/10.3390/microorganisms8030393 Psomas EI, Fletouris DJ, Litopoulou-Tzanetaki E, Tzanetakis N (2003) Assimilation of cholesterol by yeast strains isolated from infant feces and feta cheese. J Dairy Sci 86:3416–3422. https://doi.org/10.3168/jds.S0022-0302(03)73945-9 Rezac S, Kok CR, Heermann M, Hutkins R (2018) Fermented foods as a dietary source of live organisms. Front Microbiol 9:1785. https://doi.org/10.3389/fmicb.2018.01785 Rodríguez PF, Arévalo-Villena M, Rosa IZ, Perez AB (2018) Selection of potential non-saccharomyces probiotic yeasts from food origin by a step-by-step approach. Food Res Int 112:143–151. https://doi.org/10.1016/j.foodres.2018.06.008 Rosenberg M, Gutnick D, Rosenberg E (1980) Adherence of bacteria to hydrocarbons: a simple method for measuring cell-surface hydrophobicity. FEMS Microbiol Lett 9:29–33. https://doi.org/10.1111/j.1574-6968.1980.tb05599.x Rudel LL, Morris MD (1973) Determination of cholesterol using o-phthalaldehyde. J Lipid Res 14:364–366. https://doi.org/10.1016/S0022-2275(20)36896-6 Sambrani R, Abdolalizadeh J, Kohan L, Jafari B (2021) Recent advances in the application of probiotic yeasts, particularly saccharomyces, as an adjuvant therapy in the management of cancer with focus on colorectal cancer. Mol Biol Rep 48:1–10. https://doi.org/10.1007/s11033-020-06110-1 Savitri LP (2021) Probiotics for human health. In: Goel G, Kumar A (eds) Advances in probiotics for sustainable food and medicine. Microorganisms for sustainability, vol 21. Springer, Singapore. https://doi.org/10.1007/978-981-15-6795-7_8 Sen S, Mansell TJ (2020) Yeasts as probiotics: mechanisms, outcomes, and future potential. Fungal Genet Biol 137:103333. https://doi.org/10.1016/j.fgb.2020.103333 Syal P, Vohra A (2013) Probiotic potential of yeasts isolated from traditional Indian fermented foods. Int J Microbiol Res 5:390. https://doi.org/10.9735/0975-5276.5.2.390-398 Tamang JP, Watanabe K, Holzapfel WH (2016) Review: diversity of microorganisms in global fermented foods and beverages. Front Microbiol 7:377. https://doi.org/10.3389/fmicb.2016.00377 Tuo Y, Yu H, Ai L, Wu Z, Guo B, Chen W (2013) Aggregation and adhesion properties of 22 lactobacillus strains. J Dairy Sci 96:4252–4257. https://doi.org/10.3168/jds.2013-6547 Turchi B, Mancini S, Fratini F, Pedonese F, Nuvoloni R, Bertelloni F, Ebani VV, Cerri D (2013) Preliminary evaluation of probiotic potential of lactobacillus plantarum strains isolated from Italian food products. World J Microbiol Biotechnol 29:1913–1922. https://doi.org/10.1007/s11274-013-1356-7 Wulan R, Astuti RI, Rukayadi Y, Meryandini A (2021) Evaluation of indigenous Pichia kudriavzevii from cocoa fermentation for a probiotic candidate. Biodiversitas J Biolo Divers 22. https://doi.org/10.13057/biodiv/d220331 Yu Z, Zhang X, Li S, Li C, Li D, Yang Z (2013) Evaluation of probiotic properties of lactobacillus plantarum strains isolated from Chinese sauerkraut. World J Microbiol Biotechnol 29:489–498. https://doi.org/10.1007/s11274-012-1202-3 Žuntar I, Petric Z, Bursać Kovačević D, Putnik P (2020) Safety of probiotics: functional fruit beverages and nutraceuticals. Foods 9:947. https://doi.org/10.3390/foods9070947