Extracellular enzymes producing yeasts study: cost-effective production of α-amylase by a newly isolated thermophilic yeast <i>Geotrichum candidum</i> PO27

Ibtissem Chaib; Scheherazed Dakhmouche-Djekrif; Leila Bennamoun; Tahar Nouadri; Ibtissem Chaib; Scheherazed Dakhmouche-Djekrif; Leila Bennamoun; Tahar Nouadri

doi:10.3934/microbiol.2024006

AIMS Microbiology

2024, Volume 10, Issue 1: 83-106. doi: 10.3934/microbiol.2024006

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Research article

Extracellular enzymes producing yeasts study: cost-effective production of α-amylase by a newly isolated thermophilic yeast Geotrichum candidum PO27

1.
Laboratory of Microbiological Engineering and Applications, Department of Biochemistry and Molecular and Cellular Biology, Faculty of Natural and Life Sciences, Frères Mentouri University Constantine 1, Constantine 25017, Algeria
2.
Department of Natural Sciences, Teachers Training School El Katiba ‎Assia Djebar, University town Ali Mendjeli, Constantine 25000, Algeria

Received: 22 November 2023 Revised: 01 January 2024 Accepted: 22 January 2024 Published: 29 January 2024

Enzymes are biocatalysts mainly used for their industrial potential in various applications. The present study aims to understand the enzyme production for biotechnological interest from a local yeast strain. From 100 isolates obtained from various biotopes, 78 strains were selected for their enzymatic heritage. Screening of α-amylase, lipase/esterase, and cellulase activities by rapid plate detection methods was carried out and the PO27 yeast was selected for its high capacity to produce α-amylase. In addition, this yeast strain exhibited good lipolytic and esterolytic activities, as well as low cellulase activity. A sequence analysis of the D1/D2 region of the 26S ribosomal RNA (26S rRNA) and a study of morphological characteristics identified the PO27 strain as Geotrichum candidum. The production of α-amylase has been studied in solid medium fermentation using various natural substrates without any supplementation such as olive pomace, potato peels, leftover bread, and mastic cake. G. candidum PO27 showed an improved production of α-amylase with olive pomace, thus reaching approximately 180.71 U/g. To evaluate the ability of this isolate to produce α-amylase in submerged fermentation, multiple concentrations of olive pomace substrate were tested. The best activity of submerged fermentation was statistically compared to the solid-state fermentation result in order to select the appropriate fermentation type. A high significant difference was found to rank the 6% olive pomace medium as the best substrate concentration with 34.395 U/mL of α-amylase activity. This work showed that the new isolate Geotrichum candidum PO27 has a better potential to produce α-amylase at a low cost in solid-state fermentation compared to submerged fermentation. Optimization conditions for PO27 α-amylase production through solid-state fermentation were achieved ‎ using a one factor at a time (OFAT) approach. The findings revealed that a high temperature (60 °C), an acidic pH, malt extract, and soluble starch were the highly significant medium components for enhancing α-‎amylase production. The use of olive pomace waste by Geotrichum candidum PO27 is expected to be effective in producing an industrially useful α-amylase.
- Geotrichum candidum PO27,
- α-amylase,
- Olive pomace,
- SSF,
- SmF,
- OFAT
Citation: Ibtissem Chaib, Scheherazed Dakhmouche-Djekrif, Leila Bennamoun, Tahar Nouadri. Extracellular enzymes producing yeasts study: cost-effective production of α-amylase by a newly isolated thermophilic yeast Geotrichum candidum PO27[J]. AIMS Microbiology, 2024, 10(1): 83-106. doi: 10.3934/microbiol.2024006

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Abstract

Enzymes are biocatalysts mainly used for their industrial potential in various applications. The present study aims to understand the enzyme production for biotechnological interest from a local yeast strain. From 100 isolates obtained from various biotopes, 78 strains were selected for their enzymatic heritage. Screening of α-amylase, lipase/esterase, and cellulase activities by rapid plate detection methods was carried out and the PO27 yeast was selected for its high capacity to produce α-amylase. In addition, this yeast strain exhibited good lipolytic and esterolytic activities, as well as low cellulase activity. A sequence analysis of the D1/D2 region of the 26S ribosomal RNA (26S rRNA) and a study of morphological characteristics identified the PO27 strain as Geotrichum candidum. The production of α-amylase has been studied in solid medium fermentation using various natural substrates without any supplementation such as olive pomace, potato peels, leftover bread, and mastic cake. G. candidum PO27 showed an improved production of α-amylase with olive pomace, thus reaching approximately 180.71 U/g. To evaluate the ability of this isolate to produce α-amylase in submerged fermentation, multiple concentrations of olive pomace substrate were tested. The best activity of submerged fermentation was statistically compared to the solid-state fermentation result in order to select the appropriate fermentation type. A high significant difference was found to rank the 6% olive pomace medium as the best substrate concentration with 34.395 U/mL of α-amylase activity. This work showed that the new isolate Geotrichum candidum PO27 has a better potential to produce α-amylase at a low cost in solid-state fermentation compared to submerged fermentation. Optimization conditions for PO27 α-amylase production through solid-state fermentation were achieved ‎ using a one factor at a time (OFAT) approach. The findings revealed that a high temperature (60 °C), an acidic pH, malt extract, and soluble starch were the highly significant medium components for enhancing α-‎amylase production. The use of olive pomace waste by Geotrichum candidum PO27 is expected to be effective in producing an industrially useful α-amylase.

Abbreviations

SmF

submerged fermentation

SSF

state fermentation

olive pomace

olive forest soil

DNSA

dinitro salicylic acid

thermal water

mastic oil cake

vegetable smen

olive rinse water

YPSA

yeast extract peptone soluble starch agar

YPDA

yeast extract peptone dextrose agar

yeast extract malt extract

SDA

sabouraud dextrose agar

YGA

yeast extract glucose agar

YPCA

yeast extract peptone carboxymethyl cellulose agar

TPA

tween peptone agar

amylolytic index

OFAT

one factor at a time

NCBI

National Center for Biotechnology Information

Acknowledgments

The article was prepared without the use of external financial support.

Conflict of interest

All authors declare no conflicts of interest in this paper.

Author contributions

IC: methodology, experiment performing, data analysis, drafting the manuscript; SDD: conceptualization, methodology, supervised the project, editorial review; LB: formal analysis, review of the final version of the manuscript; TN: formal analysis, project administration. All authors approved the final version of the manuscript.

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