Lipase-producing Filamentous Fungi from Non-dairy Creamer Industrial Waste

Desty Triyaswati,Miftahul Ilmi 한국미생물·생명공학회 2020 한국미생물·생명공학회지 Vol.48 No.2

Lipase-producing fungi have been isolated from environments containing lipids. The non-dairy creamer industrial waste has a high amount of lipids so it is a potential source for the isolation of lipase-producing fungi. However, the study of fungi that secrete lipase from this industrial waste has not been reported. The purpose of this study was to obtain lipase-producing filamentous fungi from non-dairy creamer industrial waste. Mineral salt and potato dextrose agar were used as media for the isolation process. The qualitative screening was conducted using phenol red agar medium and the quantitative screening using broth medium containing glucose and olive oil. Isolates producing the highest amounts of lipase were identified with molecular methods. We found that 5 out of 19 isolated filamentous fungi are lipase producers. Further analysis showed that isolate Ms.11 produced the highest amount of lipase compared to others. Based on ITS sequence Ms.11 was identified as Aspergillus aculeatus. The lipase activity in medium containing 1% glucose + 1% olive oil at pH 7.0 and 30℃ after 96 and 120 h of incubation was 5.13 ± 0.30 U/ml and 5.22 ± 0.59 U/ml, respectively. The optimum lipase activity was found at pH 7.0, 30℃ and using methanol or ethanol in the reaction tube. Lipase was more stable at 20−30℃ and maintained 85% of its activity. It was concluded that isolate Ms.11 is a potential source of lipase that catalyzes transesterification reactions. Further studies are required to optimize lipase production to make the strain suitable for industry purposes.

Lipase-producing Filamentous Fungi from Non-dairy Creamer Industrial Waste

Triyaswati, Desty,Ilmi, Miftahul The Korean Society for Microbiology and Biotechnol 2020 한국미생물·생명공학회지 Vol.48 No.2

Lipase-producing fungi have been isolated from environments containing lipids. The non-dairy creamer industrial waste has a high amount of lipids so it is a potential source for the isolation of lipase-producing fungi. However, the study of fungi that secrete lipase from this industrial waste has not been reported. The purpose of this study was to obtain lipase-producing filamentous fungi from non-dairy creamer industrial waste. Mineral salt and potato dextrose agar were used as media for the isolation process. The qualitative screening was conducted using phenol red agar medium and the quantitative screening using broth medium containing glucose and olive oil. Isolates producing the highest amounts of lipase were identified with molecular methods. We found that 5 out of 19 isolated filamentous fungi are lipase producers. Further analysis showed that isolate Ms.11 produced the highest amount of lipase compared to others. Based on ITS sequence Ms.11 was identified as Aspergillus aculeatus. The lipase activity in medium containing 1% glucose + 1% olive oil at pH 7.0 and 30℃ after 96 and 120 h of incubation was 5.13 ± 0.30 U/ml and 5.22 ± 0.59 U/ml, respectively. The optimum lipase activity was found at pH 7.0, 30℃ and using methanol or ethanol in the reaction tube. Lipase was more stable at 20-30℃ and maintained 85% of its activity. It was concluded that isolate Ms.11 is a potential source of lipase that catalyzes transesterification reactions. Further studies are required to optimize lipase production to make the strain suitable for industry purposes.

Optimization of Medium for Lipase Production from Zygosaccharomyces mellis SG1.2 Using Statistical Experiment Design

( Marisa Dian Pramitasari ),( Miftahul Ilmi ) 한국미생물 · 생명공학회 2021 한국미생물·생명공학회지 Vol.49 No.3

Lipase (triacylglycerol lipase, EC 3.1.1.3) is an enzyme capable of hydrolyzing triacylglycerol, to produce fatty acids and glycerol and reverse the reaction of triacylglycerol synthesis from fatty acids and glycerol through transesterification. Applications of lipase are quite widespread in the industrial sector, including in the detergent, paper, dairy, and food industries, as well as for biodiesel synthesis. Lipases by yeasts have attracted industrial attention because of their fast production times and high stability. In a previous study, a lipase-producing yeast isolate was identified as Zygosaccharomyces mellis SG1.2 and had a productivity of 24.56 U/mg of biomass. This productivity value has the potential to be a new source of lipase, besides Yarrowia lypolitica which has been known as a lipase producer with a productivity of 0.758 U/mg. Lipase production by Z. mellis SG1.2 needs to be increased by optimizing the production medium. The aims of this study were to determine the significant component of the medium for lipase production and methods to increase lipase production using the optimum medium. The two methods used for the statistical optimization of production medium were Taguchi and RSM (Response Surface Methodology). The data obtained were analyzed using Minitab 18 and SPSS 23 software. The most significant factors which affected lipase productivity were olive oil and peptones. The optimum medium composition consisted of 1.02% olive oil, 2.19% peptone, 0.05% MgSO₄·7H₂O, 0.05% KCl, and 0.2% K₂HPO₄. The optimum medium was able to increase the lipase productivity of Z. mellis SG1.2 to 1.8-fold times the productivity before optimization.

Genetic Diversity of Amylomyces rouxii from Ragi tapai in Java Island Based on Ribosomal Regions ITS1/ITS2 and D1/D2

Delva Ega,Arisuryanti Tuty,Ilmi Miftahul 한국균학회 2022 Mycobiology Vol.50 No.2

Amylomyces rouxii is commonly found as amylolytic fungi in tapai fermentation. However, its diversity is rarely reported despite being often used for food production in Southeast Asia. This research aims to analyze the genetic diversity and the distribution pattern of A. rouxii from Ragi tapai in Java Island, Indonesia. We isolated the fungus from samples obtained from Ragi tapai producing centers in Bandung, Sumedang, Muntilan, Blora, Yogyakarta, and Bondowoso. The obtained isolates were molecularly identified based on the ribosomal regions ITS1/ITS2 and D1/D2, then analyzed for phylogenetic tree reconstruction, genetic distance, genetic variation, and haplotype networking. Six isolates showed specific morphological traits of A. rouxii. However, phylogenetic tree reconstruction on the ribosomal genes showed that the isolates were grouped into two different clades related to two species. Clade A included BDG, SMD, and MTL isolates related to A. rouxii, whereas clade B included YOG, BLR, and BDS isolates related to Mucor indicus. The genetic distances between clades for ITS1/ITS2 and D1/D2 were 0.6145 and 0.1556, respectively. In conclusion, we confirmed the genetic diversity of molds from Ragi tapai in Java Island and showed that the isolates are not only related to A. rouxii as reported before.

Potential Yeast from Indonesian Wild Forest Honey Showing Ability to Produce Lipase for Lipid Transesterification

Prayolga Toban Palilu,Rina Sri Kasiamdari,Miftahul Ilmi 한국미생물·생명공학회 2019 한국미생물·생명공학회지 Vol.47 No.4

Biodiesel is produced through the transesterification process in the presence of alcohol and a catalyst that catalyzes the conversion of triglycerides to esters and glycerol compounds. A more optimal product conversion can be achieved using enzymes, such as lipase. Lipase is reported to be produced in osmophilic yeasts due to the low water content in their natural habitats. Wild forest honey is one of the osmophilic natural habitats in Indonesia. However, lipase-producing yeast has not been reported in the Indonesian honey. In this study, we screened the lipase-producing yeasts isolated from wild forest honey collected from Central Sulawesi. The production profile and activity of lipase were determined at different pH values and temperatures. One promising yeast was isolated from the honey, which was identified as Zygosaccharomyces mellis SG 1.2 based on ITS sequence. The maximum lipase production (24.56 ± 1.30 U/mg biomass) was achieved by culturing the strain in a medium containing 2% olive oil as a carbon source at pH 7 and 30℃ for 40 h. The optimum pH and temperature for lipase activity were 6 and 55℃, respectively. The enzyme maintained 80% of its activity upon incubation at 25℃ for 4 h. However, the enzyme activity decreased by more than 50% upon incubation at 35 and 40℃ for 2 h. This is the first study to report the lipase producing capability of Z. mellis. Further studies are needed to optimize the enzyme production.

Potential Yeast from Indonesian Wild Forest Honey Showing Ability to Produce Lipase for Lipid Transesterification

Palilu, Prayolga Toban,Kasiamdari, Rina Sri,Ilmi, Miftahul The Korean Society for Microbiology and Biotechnol 2019 한국미생물·생명공학회지 Vol.47 No.4

Biodiesel is produced through the transesterification process in the presence of alcohol and a catalyst that catalyzes the conversion of triglycerides to esters and glycerol compounds. A more optimal product conversion can be achieved using enzymes, such as lipase. Lipase is reported to be produced in osmophilic yeasts due to the low water content in their natural habitats. Wild forest honey is one of the osmophilic natural habitats in Indonesia. However, lipase-producing yeast has not been reported in the Indonesian honey. In this study, we screened the lipase-producing yeasts isolated from wild forest honey collected from Central Sulawesi. The production profile and activity of lipase were determined at different pH values and temperatures. One promising yeast was isolated from the honey, which was identified as Zygosaccharomyces mellis SG 1.2 based on ITS sequence. The maximum lipase production (24.56 ± 1.30 U/mg biomass) was achieved by culturing the strain in a medium containing 2% olive oil as a carbon source at pH 7 and 30℃ for 40 h. The optimum pH and temperature for lipase activity were 6 and 55℃, respectively. The enzyme maintained 80% of its activity upon incubation at 25℃ for 4 h. However, the enzyme activity decreased by more than 50% upon incubation at 35 and 40℃ for 2 h. This is the first study to report the lipase producing capability of Z. mellis. Further studies are needed to optimize the enzyme production.