Chemical implications and time reduction of on-farm cocoa fermentation by Saccharomyces cerevisiae and Pichia kudriavzevii

This study aimed to identify the volatile compounds in the fermented and dried cocoa beans conducted with three distinct inoculants of yeast species due to their high fermentative capacity: Saccharomyces cerevisiae, Pichia kudriavzevii, the mixture in equal proportions 1:1 of both species, and a control fermentation (with no inoculum application). Three starter cultures of yeasts, previously isolated and identified in cocoa fermentation in the municipality of Tomé-Açu, Pará state, Brazil. The seeds with pulp were removed manually and placed in wooden boxes for the fermentation process that lasted from 6 to 7 days. On the last day of fermentation, the almonds were packaged properly and placed to dry (36 °C), followed by preparation for the analysis of volatile compounds by GC-MS technique. In addition to the control fermentation, a high capacity for the formation of desirable compounds in chocolate by the inoculants with P. kudriavzevii was observed, which was confirmed through multivariate analyses, classifying these almonds with the highest content of aldehydes, esters, ketones and alcohols and low concentration of off-flavours. We conclude that the addition of mixed culture starter can be an excellent alternative for cocoa producers, suggesting obtaining cocoa beans with desirable characteristics for chocolate production, as well as creating a product identity for the producing region.

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Functional Biodiversity of Yeasts Isolated from Colombian Fermented and Dry Cocoa Beans

Microorganisms ◽

10.3390/microorganisms8071086 ◽

2020 ◽

Vol 8 (7) ◽

pp. 1086 ◽

Cited By ~ 2

Author(s):

Johannes Delgado-Ospina ◽

Samantha Triboletti ◽

Valentina Alessandria ◽

Annalisa Serio ◽

Manuel Sergi ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Fermentation Process ◽

Yeast Species ◽

Degradation Products ◽

Microbial Community Composition ◽

Functional Biodiversity ◽

Cocoa Beans ◽

Pichia Kudriavzevii ◽

First Time ◽

Cocoa Fermentation

Yeasts play an important role in the cocoa fermentation process. Although the most relevant function is the degradation of sugars and the production of ethanol, there is little understanding of the enzyme activities and attributes that allow them to survive even after drying. The present study explored the functional biodiversity of yeasts associated with Criollo Colombian cocoa fermented beans, able to survive after drying. Twelve species belonging to 10 genera of osmo-, acid-, thermo-, and desiccation-tolerant yeasts were isolated and identified from fermented and dry cocoa beans, with Pichia kudriavzevii and Saccharomyces cerevisiae standing out as the most frequent. For the first time, we reported the presence of Zygosaccharomyces bisporus in cocoa fermented beans. It was found that resistance to desiccation is related to the different degradation capacities of fermentation substrates, which suggests that associative relationships may exist between the different yeast species and their degradation products. Besides, the increased thermotolerance of some species was related to the presence of polyphenols in the medium, which might play a fundamental role in shaping the microbial community composition.

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Exploring the Capability of Yeasts Isolated from Colombian Fermented Cocoa Beans to Form and Degrade Biogenic Amines in a Lab-Scale Model System for Cocoa Fermentation

Microorganisms ◽

10.3390/microorganisms9010028 ◽

2020 ◽

Vol 9 (1) ◽

pp. 28

Author(s):

Johannes Delgado-Ospina ◽

Laura Acquaticci ◽

Junior Bernardo Molina-Hernandez ◽

Kalliopi Rantsiou ◽

Maria Martuscelli ◽

...

Keyword(s):

Biogenic Amines ◽

Yeast Species ◽

Scale Model ◽

Cocoa Beans ◽

Pichia Kudriavzevii ◽

Toxic Compounds ◽

Volatile Organic ◽

Fatty Acids Metabolism ◽

Cocoa Fermentation

Yeast starters for cocoa fermentation are usually tested according to their enzymatic activities in terms of mucilage degradation and flavor improvement, disregarding their influence on the production or elimination of toxic compounds as biogenic amines (BAs), important for human health. In this work, we tested 145 strains belonging to 12 different yeast species and isolated from the Colombian fermented cocoa beans (CB) for their capability of producing BAs in vitro. Sixty-five strains were able to decarboxylate at least one of the amino acids tested. Pichia kudriavzevii ECA33 (Pk) and Saccharomyces cerevisiae 4 (Sc) were selected to evaluate their potential to modulate BAs, organic acids, and volatile organic compounds (VOCs) accumulation during a simulated cocoa fermentation. The growth of Sc or Pk in the presence of CB caused a significant reduction (p < 0.05) of 2-phenylethylamine (84% and 37%) and cadaverine (58% and 51%), and a significant increase of tryptamine and putrescine with a strong influence of temperature in BA formation and degradation. In addition, our findings pointed out that Pk induced a major production of fatty acid- and amino acid-derived VOCs, while Sc induced more VOCs derived from fatty acids metabolism. Our results suggest the importance of considering BA production in the choice of yeast starters for cocoa fermentation.

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Method-Dependent Epidemiological Cutoff Values for Detection of Triazole Resistance in Candida and Aspergillus Species for the Sensititre YeastOne Colorimetric Broth and Etest Agar Diffusion Methods

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01651-18 ◽

2018 ◽

Vol 63 (1) ◽

Cited By ~ 19

Author(s):

A. Espinel-Ingroff ◽

J. Turnidge ◽

A. Alastruey-Izquierdo ◽

F. Botterel ◽

E. Canton ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Species Complex ◽

Susceptibility Testing ◽

Yeast Species ◽

Sensu Stricto ◽

Wild Type ◽

Pichia Kudriavzevii ◽

Cutoff Value ◽

Content Type ◽

Clavispora Lusitaniae

ABSTRACT Although the Sensititre Yeast-One (SYO) and Etest methods are widely utilized, interpretive criteria are not available for triazole susceptibility testing of Candida or Aspergillus species. We collected fluconazole, itraconazole, posaconazole, and voriconazole SYO and Etest MICs from 39 laboratories representing all continents for (method/agent-dependent) 11,171 Candida albicans, 215 C. dubliniensis, 4,418 C. glabrata species complex, 157 C. guilliermondii (Meyerozyma guilliermondii), 676 C. krusei (Pichia kudriavzevii), 298 C. lusitaniae (Clavispora lusitaniae), 911 C. parapsilosis sensu stricto, 3,691 C. parapsilosis species complex, 36 C. metapsilosis, 110 C. orthopsilosis, 1,854 C. tropicalis, 244 Saccharomyces cerevisiae, 1,409 Aspergillus fumigatus, 389 A. flavus, 130 A. nidulans, 233 A. niger, and 302 A. terreus complex isolates. SYO/Etest MICs for 282 confirmed non-wild-type (non-WT) isolates were included: ERG11 (C. albicans), ERG11 and MRR1 (C. parapsilosis), cyp51A (A. fumigatus), and CDR2 and CDR1 overexpression (C. albicans and C. glabrata, respectively). Interlaboratory modal agreement was superior by SYO for yeast species and by the Etest for Aspergillus spp. Distributions fulfilling CLSI criteria for epidemiological cutoff value (ECV) definition were pooled, and we proposed SYO ECVs for S. cerevisiae and 9 yeast and 3 Aspergillus species and Etest ECVs for 5 yeast and 4 Aspergillus species. The posaconazole SYO ECV of 0.06 µg/ml for C. albicans and the Etest itraconazole ECV of 2 µg/ml for A. fumigatus were the best predictors of non-WT isolates. These findings support the need for method-dependent ECVs, as, overall, the SYO appears to perform better for susceptibility testing of yeast species and the Etest appears to perform better for susceptibility testing of Aspergillus spp. Further evaluations should be conducted with more Candida mutants.

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Potentials of multi-stress tolerant yeasts, Saccharomyces cerevisiae and Pichia kudriavzevii for fuel ethanol production from industrial cassava wastes

Process Biochemistry ◽

10.1016/j.procbio.2021.11.014 ◽

2021 ◽

Author(s):

Yoshinori Murata ◽

Charles O. Nwuche ◽

Julius E. Nweze ◽

Ifeanyi A. Ndubuisi ◽

James C. Ogbonna

Keyword(s):

Saccharomyces Cerevisiae ◽

Ethanol Production ◽

Fuel Ethanol ◽

Pichia Kudriavzevii

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Starter consortia for on-farm cocoa fermentation and their quality attributes

Preparative Biochemistry & Biotechnology ◽

10.1080/10826068.2019.1689508 ◽

2019 ◽

Vol 50 (3) ◽

pp. 272-280 ◽

Cited By ~ 2

Author(s):

Yallappa Basappa Saunshi ◽

Mudrakola Vidya Sagar Sandhya ◽

Navin Kumar Rastogi ◽

Pushpa Srinivas Murthy

Keyword(s):

Quality Attributes ◽

On Farm ◽

Cocoa Fermentation

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Genome Sequences of Cyberlindnera fabianii 65, Pichia kudriavzevii 129, and Saccharomyces cerevisiae 131 Isolated from Fermented Masau Fruits in Zimbabwe

Genome Announcements ◽

10.1128/genomea.00064-17 ◽

2017 ◽

Vol 5 (14) ◽

Cited By ~ 6

Author(s):

Irma M. H. van Rijswijck ◽

Martijn F. L. Derks ◽

Tjakko Abee ◽

Dick de Ridder ◽

Eddy J. Smid

Keyword(s):

Saccharomyces Cerevisiae ◽

Genome Sequences ◽

Pichia Kudriavzevii ◽

Content Type ◽

Food Applications

ABSTRACT Cyberlindnera fabianii 65, Pichia kudriavzevii 129, and Saccharomyces cerevisiae 131 have been isolated from the microbiota of fermented masau fruits. C. fabianii and P. kudriavzevii especially harbor promising features for biotechnology and food applications. Here, we present the draft annotated genome sequences of these isolates.

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Isolation and characterization of yeast from different sources

Mongolian Journal of Agricultural Sciences ◽

10.5564/mjas.v29i1.1374 ◽

2020 ◽

Vol 29 (1) ◽

pp. 86-92

Author(s):

Munkh-Erdene Sugarragchaa ◽

Lhagvadolgor Batjargal ◽

Narangerel Mijid ◽

Urantulkhuur Battumur

Keyword(s):

Saccharomyces Cerevisiae ◽

Dna Sequence ◽

Kluyveromyces Marxianus ◽

Genomic Dna ◽

Evolutionary Relationship ◽

Candida Boidinii ◽

Rrna Gene ◽

Pichia Kudriavzevii ◽

Isolation And Characterization ◽

Genomic Dna Sequence

Four strains of yeasts isolated from some mare’s fermented milk, whey and fruit were identified using their morphology, biochemistry and phylogenetic characteristics. These yeasts belonged to four genera viz: Candida (one strain), Pichia (one strain), Saccharomyces (one strain) and Kluyveromyces (one strain). The physiological and biochemical tests of the yeasts carried out showed all isolates to ferment glucose for their growth. All isolates showed elliptical to round spores. Pichia kudriavzevii was able to ferment xylose of the four isolates. The assimilation and fermentation of most sugars by the isolates was variable. The blast sequence query showed that Saccharomyces cerevisiae and Pichia kudriavzevii had maximum identity (100%) with the genomic DNA sequence of MON-21 and MON-22, respectively at both ITS. Isolate Candida boidinii MON-23 was 100% homologous to Candida boidinii 18S rRNA gene sequence. The blast sequence query showed that Kluyveromyces marxianus MON-24 has the maximum identity (99%) with the genomic DNA sequence of Kluyveromyces marxianus at ITS 1 and ITS 4 sequence with that in the Genbank Library Database. Using MEGA 7 software, the phylogenetic trees were constructed to determine the evolutionary relationship of newly identified yeasts from our experiment and previously published yeast species. The sequences of the yeasts were deposited in the National Center for Biotechnology Information (NCBI) database. Ялгаатай эх үүсвэрээс ялгасан хөрөнгөнцрийн шинж чанар Монголчуудын эрт үеэс хэрэглэж ирсэн зарим уламжлалт исгэлэн бүтээгдэхүүн, сүү, сүүн бүтээгдэхүүний дайвар түүхий эд болох шар сүү, жимснээс хөрөнгөнцрийн цэвэр өсгөвөр ялган авч, тэдгээрийн морфологи, биохими, филогенетикийн шинж чанарыг судлав. МОН-21, 22, 23 болон 24 өсгөвөрүүд нь грам эерэг, эсийн урт нь 1.2-10.8 мкм, эсийн өргөн нь 0.5-6.0 мкм хэмжээтэй, пигмент үүсэлт нь цайвар шар өнгөтэй, жигд бөөгнөрсөн, дугуй, товгор хэлбэртэй, МОН-21 өсгөвөр нь нүүрстөрөгчийн эх үүсвэрээр галактоз, декстроз, МОН-22 өсгөвөр нь ксилоз декстроз, МОН-23 өсгөвөр нь сахароз, декстроз, ксилоз, фрюктоз, МОН-24 өсгөвөр нь мальтоз, ксилозыг тус тус ашиглаж чадахгүй байна. Дрожжийн ангилал зүйг ITS ген ашиглан тодорхойлоход, МОН-21 өсгөвөр нь Pichia kudriavzevii DGY49-тай 100%, МОН-22 нь Saccharomyces cerevisiae S5-тай 100%, МОН-23 өсгөвөр нь Candida boidinii NRRL Y-2332-тай 99.88%, МОН-24 өсгөвөр нь Kluyveromyces marxianus CCT 7735-тай 99.88% зүйлд тус тус ижил төсөөтэй байна. Мөн ITS1 болон ITS4 праймеруудыг ашиглан ялгасан өсгөвөрүүдийн зүйлийн генетик хамаарлыг тодорхойлоход, өсгөвөр МОН-23 болон МОН-24 нь бусад өсгөврүүдээс өөр хэмжээтэй буюу жижиг 18S рРНХ-ийн генийн молекултай, МОН-21 болон МОН-22 өсгөвөрүүд нь том 28S рРНХ-ийн генийн молекул хэмжээтэй байна. Эдгээр өсгөвөрүүдийг цаашид нарийвчлан судалж, исгэлтийн үйлдвэрлэлд ашиглах боломжтой хөрөнгө гарган авах, цэвэр өсгөврийн сан байгуулах, бүртгэлжүүлэх, дата мэдээлэл үүсгэх, үндэсний үйлдвэрлэлийг хөрөнгөөр хангах боломжийг нээж өгнө. Түлхүүр үг: хөрөнгөнцөр, kluyveromyces marxianus, рРНХ, pichia kudriavzevii, декстроз

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Isolation and identification of microbial species found in cocoa fermentation as microbial starter culture candidates for cocoa bean fermentation in Colombia.

Pelita Perkebunan (a Coffee and Cocoa Research Journal) ◽

10.22302/iccri.jur.pelitaperkebunan.v36i3.443 ◽

2020 ◽

Vol 36 (3) ◽

pp. 236-248

Author(s):

Maria Denis Lozano Tovar ◽

Geraldine Tibasosa ◽

Carlos Mario González ◽

Karen Ballestas Alvarez ◽

Martha Del Pilar Lopez Hernandez ◽

...

Keyword(s):

Acetic Acid ◽

Starter Culture ◽

Cocoa Bean ◽

Rrna Gene ◽

Cocoa Beans ◽

Isolation And Identification ◽

Pichia Kudriavzevii ◽

Cocoa Bean Fermentation ◽

Cocoa Fermentation

Microbial activity involved in the cocoa beans fermentation process is essential to maintain and improve the organoleptic and nutritional qualities of chocolate; therefore, the aim of this investigation was to search and select microbial isolates with the potential to improve the quality of cocoa beans. Fermentation experimentswere conducted on farms located in Maceo (Antioquia), San Vicente de Chucurí (Santander), and Rivera and Algeciras (Huila), Colombia. Yeast, lactic acid bacteria (LAB), acetic acid bacteria (AAB), and mesophilic aerobic microorganisms were obtained from different fermentation batches. The growth of these microorganismswas tested in six treatments as follows: 50% cocoa pulp agar (CPA), high concentrations of glucose (10%), ethanol (5%), and acetic acid (7%), an acidic pH of 3.0, and a high temperature of 50oC for 24 h. The isolates with the highest growth were identified by 18S and 16S rRNA gene analysis, revealing a high diversity ofspecies associated with cocoa fermentation, including eight species of yeasts (Debaryomyces hansenii, Meyerozyma guillermondii, Wickerhanomyces anomalus, Pichia guillermondii, Pichia kudriavzevii, Trichosporon asahii, Candida parapsilosis, and Pichia manshurica), six species of LAB (Pediococcus acidilactici, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus farraginis, Lactobacillus rhamnosus, and Leuconostoc mesenteroides), four species of AAB (Gluconobacter japonicus, Acetobacter tropicalis, Acetobacter pasteurianus, and Acetobacter malorum/tropicalis), and three species of Bacillus spp. (Bacillusaryabhattai /megaterium, Bacillus subtilis, and Bacillus coagulans). In general, microbial populations increased in cocoa batches after 12 h of fermentation and decreased after 84-96 h. All the yeast isolates grew in 10% glucose and CPA, 85.7% in 5% ethanol, and 95% at a pH of 3.0. All the yeast isolates were affectedby 7% acetic acid and incubation at 50oC for 24 h. Eighty-five percent of the LAB grew in 10% glucose, 100% in 5% ethanol, 42.8% in CPA, 64% at a pH of 3.0, and 35.7% grew after being exposed to 50oC for 24 h; all were affected by 7% acetic acid. As for the AAB, 100% grew in 10% glucose, 71% in 7% ethanol, 100% grew in CPA, in 7% acetic acid, and at a pH of 3.0, while 100% were affected by incubation at 50oC. Three yeast isolates, W. anomalus, D. hansenii and M. guillermondii, three LAB isolates, P. acidilactici, L. brevis, and L. plantarum, and three AAB isolates, A. tropicalis, A. pasteurianus and G. japonicus, were selected as promising strains to be used in a microbial starter culture for cocoa bean fermentation to improve the organoleptic quality of cocoa.

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