Agroindustrial Wastes as Alternative for Lipase Production by Candida viswanathii under Solid-State Cultivation: Purification, Biochemical Properties, and Its Potential for Poultry Fat Hydrolysis

The aims of this work were to establish improved conditions for lipase production by Candida viswanathii using agroindustrial wastes in solid-state cultivation and to purify and evaluate the application of this enzyme for poultry fat hydrolysis. Mixed wheat bran plus spent barley grain (1 : 1, w/w) supplemented with 25.0% (w/w) olive oil increased the lipase production to 322.4%, compared to the initial conditions. When olive oil was replaced by poultry fat, the highest lipase production found at 40% (w/w) was 31.43 U/gds. By selecting, yeast extract supplementation (3.5%, w/w), cultivation temperature (30°C), and substrate moisture (40%, w/v), lipase production reached 157.33 U/gds. Lipase was purified by hydrophobic interaction chromatography, presenting a molecular weight of 18.5 kDa as determined by SDS-PAGE. The crude and purified enzyme showed optimum activity at pH 5.0 and 50°C and at pH 5.5 and 45°C, respectively. The estimated half-life at 50°C was of 23.5 h for crude lipase and 6.7 h at 40°C for purified lipase. Lipase presented high activity and stability in many organic solvents. Poultry fat hydrolysis was maximum at pH 4.0, reaching initial hydrolysis rate of 33.17 mmol/L/min. Thus, C. viswanathii lipase can be successfully produced by an economic and sustainable process and advantageously applied for poultry fat hydrolysis without an additional acidification step to recover the released fatty acids.

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Use of agro-industrial residues for lipase production by Candida viswanathii in a solid-state cultivation system

Scientia Plena ◽

10.14808/sci.plena.2021.081513 ◽

2021 ◽

Vol 17 (8) ◽

Author(s):

Alanna Cristinne Martins Lima ◽

Camila Moitinho Dos Santos ◽

Iara Leandro Dos Santos ◽

Lunara Thaís Alves de Bastos ◽

Annanda Carvalho dos Santos ◽

...

Keyword(s):

Solid State ◽

Olive Oil ◽

Yeast Extract ◽

Seed Coat ◽

Scale Up ◽

Soybean Seed ◽

Lipase Production ◽

Poultry Fat ◽

Candida Viswanathii ◽

Soybean Seed Coat

Lipases are an important group of enzymes that catalyze the hydrolysis of triacylglycerol and there are many industrial applications. The aim of this work was to produce the lipase by the yeast Candida viswanathii using solid state culture with agro-industrial wastes (barley bagasse, corn husk, corncob, soybean seed coat and soybean husk). The biomass pretreatment methods were evaluated, as well as the media supplementation with nitrogen and mixing substrates. Also, the efficiency of olive oil and poultry fat was evaluated on the induction of lipase production, followed by the scale-up from 20 g to 100 g. The enzyme activities in the cultures without pretreatement were higher when soybean seed coat supplemented with both olive oil (7.06 U/gss) and poultry fat (8.40 U/gss) were used. However, the pretreated substrates did not demonstrate a satisfying induction of lipolytic activity. From the nitrogen sources, yeast extract showed an increase of approximately twice the original production with both olive oil (18.12 U/gss) and poultry fat (15.98 U/gss) supplementation. On the scale-up step, the results demonstrated that, for the 20 g culture, the best lipase production was observed on the 7th day (33.52 U/gss), while for the 100 g culture the highest lipase activity was after 5 days (17.88 U/gss). The cultivation of ground soybean skin without pretreatment supplemented with yeast extract as a source of nitrogen, with fresh barley bagasse and poultry fat was the best combination.

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Optimization of Lipase Production in Solid-State Fermentation by Rhizopus Arrhizus in Nutrient Medium Containing Agroindustrial Wastes

The Open Biotechnology Journal ◽

10.2174/1874070701812010189 ◽

2018 ◽

Vol 12 (1) ◽

pp. 189-203 ◽

Cited By ~ 1

Author(s):

Georgi Dobrev ◽

Hristina Strinska ◽

Anelia Hambarliiska ◽

Boriana Zhekova ◽

Valentina Dobreva

Keyword(s):

Solid State ◽

Moisture Content ◽

Solid State Fermentation ◽

Lipase Activity ◽

Nutrient Medium ◽

Solid Medium ◽

Low Cost ◽

Lipase Production ◽

Rhizopus Arrhizus ◽

Agroindustrial Wastes

Background: Rhizopus arrhizus is a potential microorganism for lipase production. Solid-state fermentation is used for microbial biosynthesis of enzymes, due to advantages, such as high productivity, utilization of abundant and low-cost raw materials, and production of enzymes with different catalytic properties. Objective: The objective of the research is optimization of the conditions for lipase production in solid-state fermentation by Rhizopus arrhizus in a nutrient medium, containing agroindustrial wastes. Method: Biosynthesis of lipase in solid-state fermentation by Rhizopus arrhizus was investigated. The effect of different solid substrates, additional carbon and nitrogen source, particles size and moisture content of the medium on enzyme production was studied. Response surface methodology was applied for determination of the optimal values of moisture content and tryptone concentration. A procedure for efficient lipase extraction from the fermented solids was developed. Results: Highest lipase activity was achieved when wheat bran was used as a solid substrate. The addition of 1% (w/w) glucose and 5% (w/w) tryptone to the solid medium significantly increased lipase activity. The structure of the solid medium including particles size and moisture content significantly influenced lipase production. A mathematical model for the effect of moisture content and tryptone concentration on lipase activity was developed. Highest enzyme activity was achieved at 66% moisture and 5% (w/w) tryptone. The addition of the non-ionic surfactant Disponyl NP 3070 in the eluent for enzyme extraction from the fermented solids increased lipase activity about three folds. Conclusion: After optimization of the solid-state fermentation the achieved 1021.80 U/g lipase activity from Rhizopus arrhizus was higher and comparable with the activity of lipases, produced by other fungal strains. The optimization of the conditions and the use of low cost components in solid-state fermentation makes the process economicaly effective for production of lipase from the investigated strain Rhizopus arrhizus.

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LIP4 gene expression from Candida viswanathii strain

10.21203/rs.3.rs-27743/v1 ◽

2020 ◽

Author(s):

Ronan C. Teixeira ◽

Matheus M. Daude ◽

Nayra M. L. de Oliveira ◽

Alex Fernando Almeida ◽

Horllys G. Barreto

Keyword(s):

Gene Expression ◽

Olive Oil ◽

Culture Media ◽

Carbon Sources ◽

In Silico Analysis ◽

Lipase Production ◽

Submerged Cultivation ◽

Degenerate Primers ◽

Candida Viswanathii ◽

High Level

Abstract Background : Lipases are hydrolases that catalyze the cleavage of triglyceride esters bonds, releasing glycerol and free fatty acids. Candida genus can produce distinct isoforms of lipase, among of them, Candida viswanathii strain is a potential lipase producer using hydrophobic carbons sources in which produced high level of enzyme under submerged cultivation using olive oil as carbon sources. This enzyme has commercially attractive due to characteristics desired in the industries processes. The genes responsible for encoding the lipases comprise a family called LIP gene. C. viswanathii not have its genome sequenced and are not available in annotated form through the GenBank nucleotide sequence database for lipase production. The aim of this work was understanding at molecular level the effect of carbon sources to lipase production, to identify and to analyses the gene expression of CvLIP4 from C. viswanathii on different culture media. Results : In silico analysis was carried out with LIP4 gene from Candida species. Degenerate primers were designed and evaluated for expression in different conditions. CvLIP4 expression was evaluated using carbon sources glucose, tributyrin, triolein and olive oil. Triolein and olive oil were strong inducer for CvLIP4 gene expression, while tributyrin was a weak inducer and glucose was strong repressor. Conclusions : These results will contribute to further studies about regulation of the lipase genes expression from C. viswanathii and heterologous expression of this enzyme to improve the catalytic conditions in industries processes.

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Lipase production by Yarrowia lipolytica using olive oil processing wastes as substrates

Journal of the Serbian Chemical Society ◽

10.2298/jsc120905005m ◽

2013 ◽

Vol 78 (6) ◽

pp. 781-794 ◽

Cited By ~ 14

Author(s):

Omar Moftah ◽

Sanja Grbavcic ◽

Walid Moftah ◽

Nevena Lukovic ◽

Olivera Prodanovic ◽

...

Keyword(s):

Solid State ◽

Olive Oil ◽

Yarrowia Lipolytica ◽

Solid State Fermentation ◽

Lipolytic Activity ◽

Lipase Production ◽

Olive Mill Wastewater ◽

Submerged Cultivation ◽

Oil Processing ◽

Olive Mill

In this study solid and liquid wastes from the olive oil processing industry were evaluated as substrates for Yarrowia lipolytica growth with the aim of lipase production. Olive mill wastewater and olive oil cake seemed to provide necessary nutrients and physical support for the yeast to grow and produce enzyme. The highest lipolytic activity of 850 IU dm-3 was achieved after 4 days of submerged cultivation in supplemented olive mill wastewater. In addition, olive oil cake appeared to be a convenient substrate for lipase production under solid state fermentation mode. Lipase production was further improved by media supplementation and/or change in physical settings of the experiment. However, the most significant improvement of lipase production under solid state fermentation was achieved by an alkaline treatment of the substrate (more than 10-fold) when the amount of produced lipase reached up to ~40 IU g-1 of substrate.

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