Optimization of Bioprocess Variables for Fungal Lipase Production using Statistical Experimental Design: A Mini Review

Lipases hold the important role in wide spectrum of biotechnological applications. Lipases are unique, not only due to their ability to perform hydrolysis of fats into fatty acids and glycerol at the water-lipid interface, but also can reverse the reaction in non-aqueous media. Currently, the studies revealed that fungi are the one of choice for lipase production. Some important lipase-producing fungal genera include Aspergillus, Penicillium, Rhizopus, Candida, Geotrichum, etc. The development of response surface methodology applied to optimize fungal lipase production is discussed.

Proteases from Latex of Euphorbia spp. and Its Application on Milk Clot Formation

Crude proteases were extracted from Euphorbiaceae family, i.e. E. milii var imperata, E. trigona, and E. maculata. Among those three crude proteases, the activity of protease from E. trigona was the highest (812.50 U/ml), whereas E. milii and E. maculata crude proteases activity were 298.60 U/ml and 95.80 U/ml, respectively. E. maculata protein concentration was the highest among those three crude enzymes (1.206 mg/ml). The optimum pH and temperature of the enzymes were pH 7.0, pH 6.0, pH 6.5 and 60 °C, 50 °C, and 50 °C, respectively. Crude protease from E. milii var imperata, E. trigona, and E. maculata retained proteolytic activity over a wide range of pH (5.0–9.0) and temperature (up to 65 °C) with casein as substrate. All crude proteases showed milk clotting activity ranged from 0.58 U/ml to 1.01 U/ml. Thus, these crude proteases are potential to be applied in dairy industries. However, further study on enzyme purification and characterization are necessary to obtain high purity of proteases before its application.Protease kasar berhasil diekstrak dari tanaman family Euphorbiaceae, yaitu E. milii var imperata, E. trigona, dan E. maculata. Diantara ketiga protease tersebut, aktivitas protease tertinggi diperoleh dari E. trigona (812,50 U/ml), sedangkan aktivitas protease dari E. milii dan E. maculata adalah 298,60 U/ml dan 95,80 U/ml, berturut-turut. Konsentrasi total protein tertinggi terdapat pada protease kasar E. maculata (1,206 mg/ml). pH dan suhu optimum ketiga enzim tersebut adalah pH 7.0, pH 6.0, pH 6.5 dan suhu 60 °C, 50 °C, and 50 °C, berturut-turut. Protease kasar dari E. milii var imperata, E. trigona, dan E. maculata menunjukkan aktivitas proteolitik pada rentang pH 5.0–9.0 dan rentang suhu sampai 65 °C menggunakan kasein sebagai substrat. Semua protease kasar menunjukkan aktivitas penggumpalan susu dengan rentang dari 0,58 U/ml sampai 1,01 U/ml. Berdasarkan hasil yang diperoleh, protease kasar dari ketiga jenis tanaman ini berpotensi untuk diaplikasikan dalam industri olahan susu. Meskipun demikian, studi lanjut mengenai purifikasi dan karakterisasi sangat diperlukan untuk memperoleh protease murni sebelum aplikasi dalam industri makanan, khususnya pada industri olahan susu.</p

Statistical Optimization for Cost-Effective Production of Yeast-Bacterium Cell-Bound Lipases Using Blended Oily Wastes and Their Potential Applications in Biodiesel Synthesis and Wastewater Bioremediation

Oily wastes have been widely used to produce lipases, but there is insufficient knowledge on their use to efficiently produce cell-bound lipases (CBLs). This research aimed to optimize yeast&ndash;bacterium CBLs production using blended oily wastes by statistical optimization and their potential applications in biodiesel production and wastewater bioremediation. The co-culture of Magnusiomyces spicifer AW2 and Staphylococcus hominis AUP19 produced CBLs as high as 4709 U/L with cell biomass of 23.4 g/L in a two-fold diluted palm oil mill effluent (POME) added by 2.08% (v/v) waste frying oil, 1.72.0% (w/v) ammonium sulfate, 0.1% (w/v) Gum Arabic as an emulsifier (initial pH at 7.0) within 24 h. The CBLs were successfully applied as whole-cell biocatalysts to produce biodiesel through esterification and transesterification with 76% and 87% yields, respectively. Direct application of CBLs for bioremediation of heat-treated various POME concentrations achieved 73.3% oil and grease removal and 73.6% COD removal within 3 days. This study has shown that the blended oily wastes medium was suitable for low-cost production of yeast&ndash;bacterium CBLs and their potential applications in solvent-free biodiesel production and wastewater bioremediation. These strategies may greatly contribute to economical green biofuel production and waste biotreatment

Statistical Optimization for Cost-Effective Production of Yeast-Bacterium Cell-Bound Lipases Using Blended Oily Wastes and Their Potential Applications in Biodiesel Synthesis and Wastewater Bioremediation

Oily wastes have been widely used to produce lipases, but there is insufficient knowledge on their use to efficiently produce cell-bound lipases (CBLs). This research aimed to optimize yeast–bacterium CBLs production using blended oily wastes by statistical optimization and their potential applications in biodiesel production and wastewater bioremediation. The co-culture of Magnusiomyces spicifer AW2 and Staphylococcus hominis AUP19 produced CBLs as high as 4709 U/L with cell biomass of 23.4 g/L in a two-fold diluted palm oil mill effluent (POME) added by 2.08% (v/v) waste frying oil, 1.72.0% (w/v) ammonium sulfate, 0.1% (w/v) Gum Arabic as an emulsifier (initial pH at 7.0) within 24 h. The CBLs were successfully applied as whole-cell biocatalysts to produce biodiesel through esterification and transesterification with 76% and 87% yields, respectively. Direct application of CBLs for bioremediation of heat-treated various POME concentrations achieved 73.3% oil and grease removal and 73.6% COD removal within 3 days. This study has shown that the blended oily wastes medium was suitable for low-cost production of yeast–bacterium CBLs and their potential applications in solvent-free biodiesel production and wastewater bioremediation. These strategies may greatly contribute to economical green biofuel production and waste biotreatment

Effect of co-substrate on production of poly-β- hydroxybutyrate (PHB) and copolymer PHBV from newly identified mutant Rhodobacter sphaeroides U7 cultivated under aerobic-dark condition

Photosynthetic bacterial mutant strain U7 was identified using both classical and molecular (16S rDNA) techniques to be Rhodobacter sphaeroides. The glutamate-acetate (GA) medium containing sodium acetate and sodium glutamate as carbon and nitrogen sources was used for production of poly-β-hydroxybutyrate (PHB) from R. sphaeroides U7 cultivated under aerobic-dark condition (200 rpm) at 37oC. Effect of auxiliary carbon sources (propionate and valerate) and concentrations (molar ratio of 40/0, 40/20, 40/40 and 40/80) on copolymer production were studied. Both combinations of acetate with valerate and acetate with propionate were found to induce the accumulation of poly-β-hydroxybutyrate-co-β-hydroxyvalerate (PHBV) within the cell. Acetate with propionate in the molar ratio of 40/40 gave the highest poly-β-hydroxyalkanoates (PHA) content (77.68%), followed by acetate with valerate at the same molar ratio (77.42%). Although their polymer contents were similar, the presence of 40 mM valerate gave more than 4 times higher hydroxyvalerate (HV) fraction (84.77%) than in the presence of 40 mM propionate (19.12% HV fraction)

Bio-Succinic Acid Production from Palm Oil Mill Effluent Using <i>Enterococcus gallinarum</i> with Sequential Purification of Biogas

Bio-succinic acid production using microorganisms has been interesting as an environmentally friendly process. Palm oil mill effluent (POME) was considered as a cheap substrate to lower the cost of production. It was revealed that 2-fold diluted POME produced more succinic acid than undiluted and 5-fold diluted POME. In addition, the effects of various neutralizing agents on succinic acid production utilized to manage pH and CO2 supply indicated that the utilization of MgCO3 as a neutralizing agent produced succinic acid of 11.5 g/L with a small amount of by-product synthesis. Plackett–Burman Design (PBD) was used to screen the most significant nutrients for bio-succinic acid production from 2-fold diluted POME using E. gallinarum. From the Pareto chart, MgCO3 and peptone presented the highest positive effect on the production of succinic acid. In addition, Box–Behnken Design (BBD) was conducted to increase bio-succinic acid production. Experiments showed the highest production of succinic acid of 23.7 g/L with the addition of 22.5 g/L MgCO3 and 12.0 g/L peptone in 2-fold diluted POME. Moreover, the experiment of replacing MgCO3 with CO2 from biogas resulted in 19.1 g/L of succinic acid, simultaneously creating the high purity of biogas and a higher CH4 content