Proteolytic activities of bacteria, yeasts and filamentous fungi isolated from coffee fruit (Coffea arabica L.)

Rodarte, Mírian Pereira; Dias, Disney Ribeiro; Vilela, Danielle Marques; Schwan, Rosane Freitas

doi:10.4025/actasciagron.v33i3.6734

Cited by 31 publications

(5 citation statements)

References 30 publications

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“…protease determined at 60.34 U/mL in this study was previously reported as 40 mg/mL in the isolate in 2017 (24) to 157.50 U/mg (25). In this study, the Klebsiella aerogenes protease was 33.27 U/mL, previously detected at 14.24 U/mL (26). Similar differences were observed in Stenotrophomonasmaltophilia protease activity which was 57.59 U/mL in this study; nevertheless, it was previously determined at 1.87 U/mg (27) to 418 U/mL (28).…”

Section: Discussionsupporting

confidence: 65%

Replacement of Trypsin by Proteases for Medical Applications

et al. 2022

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Background: Cell culture has a crucial role in many applications in biotechnology. The production of vaccines, recombinant proteins, tissue engineering, and stem cell therapy all need cell culture. Most of these activities needed adherent cells to move, which should be trypsinized several times until received on a large scale. Although trypsin is manufactured from the bovine or porcine pancreas, the problem of contamination by unwanted animal proteins, unwanted immune reactions, or contamination to pathogen reagents is the main problem. Objectives: This study investigated microbial proteases as a safe alternative for trypsin replacement in cell culture experiments for the detachment of adherent cells. Methods: The bacteria were isolated from the leather industry effluent based on their protease enzymes. After sequencing their 16S ribosomal deoxyribonucleic acid, their protease enzymes were purified, and their enzyme activities were assayed. The alteration of enzymatic activities using different substrates and the effect of substrate concentrations on enzyme activities were determined. The purified proteases were evaluated for cell detachment in the L929 fibroblast cells compared to trypsin. The separated cells were cultured again, and cell proliferation was determined by the MTT assay. Results: The results showed that the isolated bacteria were Bacillus pumilus, Stenotrophomonas sp., Klebsiella aerogenes, Stenotrophomonas maltophilia, and Bacillus licheniformis. Among the isolated bacteria, the highest and the lowest protease activity belonged to Stenotrophomonas sp. and K. aerogenes, with 60.34 and 11.09 U/mL protease activity, respectively. All the isolated microbial proteases successfully affected L929 fibroblast cells’ surface proteins and detached the cells. A significant induction in cell proliferation was observed in the cells treated with K. aerogenes protease and B. pumilus protease, respectively (P < 0.05). Conclusions: The obtained results suggested that microbial proteases can be used as safe and efficient alternatives to trypsin in cell culture in biopharmaceutical applications.

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Section: Discussionsupporting

confidence: 65%

Replacement of Trypsin by Proteases for Medical Applications

et al. 2022

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“…Protease activity was measured as follows: 2.5 mL of yeast culture (~10 8 cells/mL) was added to an Erlenmeyer flask containing 100 mL of YCB medium supplemented with 0.01% sodium caseinate and 0.1% glucose. Flasks were then incubated with agitation (90× g ) at 28°C for 3, 7, and 14 days and growth was determined from the optical density (OD) measurements at 600 nm [ 20 ].…”

Section: Methodsmentioning

confidence: 99%

Mycological evaluation of frozen meat with special reference to yeasts

et al. 2023

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Background and Aim: Fungi can play beneficial and detrimental roles in meat products; however, the diversity and significance of fungi in meat products are poorly understood. This study aimed to isolate and characterize fungal species from frozen beef samples collected from retail stores in the Qena Governorate, Egypt. Materials and Methods: A total of 70 frozen beef samples were collected from retail stores in Qena, Egypt. All samples were subjected to mycological examination. Fungal colonies were identified using conventional approaches, as well as the VITEK 2 system and DNA sequencing of the internal transcribed spacer region. Analyses of enzymatic activity, biofilm formation ability, and the antimicrobial resistance profiles of the isolated yeasts were also conducted. Results: Molds and yeasts were isolated from 40% and 60% of meat samples, respectively. Mold isolates were dominated by Aspergillus, Penicillium, and Cladosporium spp., whereas yeast isolates were identified as Candida albicans, Candida parapsilosis, Yarrowia lipolytica, Saccharomyces cerevisiae, and Rhodotorula mucilaginosa. Compared to other yeast species, the highest production of lipase and protease was observed in Candida species. The strongest ability to form biofilms was observed in Candida spp., followed by S. cerevisiae, Y. lipolytica, and R. mucilaginosa. The results of antimicrobial susceptibility testing revealed that all yeast isolates showed notable resistance to fluconazole and itraconazole. Conclusion: A significant correlation between antimicrobial resistance and biofilm formation was observed in several species. This study highlights the importance of the dangers of yeasts in food products and the extent of their impact on public health. Keywords: antimicrobial resistance, beef, biofilm, enzymatic activities, fungi, meat, yeast.

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“…The pure cultures were stored in an ultra-freezer at -80 °C in the same broth culture media used for plating, containing 20% glycerol (w/w). Yeast colonies were characterized for morphology and biochemical assessments as described by [43]. Filamentous fungi were observed with an optical microscope for preliminary identi cation.…”

Section: Methodsmentioning

confidence: 99%

Physicochemical and microbiological dynamics in composting of organic waste using a new bioreactor model

Gaspar

Assis

Carvalho

et al. 2020

Preprint

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Background It is important to use renewable resources to minimize the environmental risks and the composting is one of the most sustainable methods for the management of organic waste. Methods The objective of this work was to evaluate the physicochemical dynamics of microorganisms and to study cultivable microorganisms during the composting process of organic residues in a new model of bioreactor. The formulation of a possible cocktail of microorganisms selected for use as a compound accelerator will be further investigated. In addition, the use of two inoculants (non-commercial inoculum (NCI)) and commercial inoculum (CI)) and a control without inoculant during the composting process will be analyzed to evaluate its efficiency. Composting was performed by mixing organic waste from the garden waste and University Restaurant, obtaining an ideal C/N ratio of 30:1. The composting process was carried out in 1 m3 composters with controlled temperature and aeration. Results The thermophilic phase for all treatments was reached by the second day. Mature compost was obtained after an average of 120 days and composting in all treatments showed an increase in the availability of P and micronutrients. During composting, the population of bacteria and actinobacteria were higher than those of yeasts and filamentous fungi. Conclusions Increased composting efficiency was observed when starter cultures were used, the treatments presented advantages such as greater mineralization of P-available and micronutrients as Mn and Zn, in terms of the quality of the final product in comparison to the control treatment.

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Proteolytic activities of bacteria, yeasts and filamentous fungi isolated from coffee fruit (Coffea arabica L.)

Cited by 31 publications

References 30 publications

Replacement of Trypsin by Proteases for Medical Applications

Replacement of Trypsin by Proteases for Medical Applications

Mycological evaluation of frozen meat with special reference to yeasts

Physicochemical and microbiological dynamics in composting of organic waste using a new bioreactor model

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