Puspadewi et al. Int J App Pharm, Vol 13, Special Issue 3, 2021, 62-65 4 th International Seminar on Pharmaceutical Science and Technology 2020 | 62 OPTIMIZATION OF LACTOBACILLUS PLANTARUM ACTIVITIES IN THE BIOSINTHESIS OF LIPASE ENZYMES Original Article RIRIN PUSPADEWI * , PUTRANTI ADIRESTUTI, MIRA A. DEWI, INNANI MUKARROMAH, YUNIAR RAHMADINNI Faculty of Pharmacy, Jenderal Achmad Yani University, Jl. Terusan Jenderal Sudirman, Cimahi, West Java 40531, Indonesia Email: ririn.puspadewi@lecture.unjani.ac.id Received: 29 Sep 2020, Revised and Accepted: 20 Oct 2020 ABSTRACT Objective: Lipase was protein compounds that can be used for many human activities. Its main function was to degrade fat including 'wrapping' cholesterol which make easily flowed in the blood. The presence lipase was important because can help the digestive healthy. These enzyme can catalyze a variety of reactions including hydrolysis, alcoholysis, esterification and aminolysis. Lipase was utilized in various sectors, such as fat, oil, milk and pharmaceutical industries. This enzyme biosynthesis can be carried out by Pseudomonas aeruginosa, Lactobacillus plantarum and Aspergillus niger. Methods: The process through fermentation techniques in lipid containing substrates under optimal conditions required by microorganisms. The fermentation products produced were tested for the presence of lipase enzymes qualitatively and quantitatively. The biosynthesis process can be influence by changes in pH, temperature and the presence of glucose. This study aimed to determine the ability of L. plantarum to produce lipases with vegetables oil substrates. The research used L. plantarum carried out at 37 °C for 24-48 h and pH 6-8 in the vegetable oil substrates. Results: The fermentation products showed hydrolysis reaction to the test media containing oil lipid with lipase levels of 2.708-3.3000 U/ml Conclusion: The results showed that Lactobacillus plantarum can synthesize the lipase enzyme in palm oil and corn oil as substrates. Keywords: Lactobacillus plantarum, Biosynthesis, Lipase, Palm oil, Corn oil © 2021 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/) DOI: https://dx.doi.org/10.22159/ijap.2021.v13s3.13 Journal homepage: https://innovareacademics.in/journals/index.php/ijap INTRODUCTION Lactobacillus plantarum is a broad member of the genus Lactobacillus. It is one of the most studied species that is used in the food industry as a probiotic microorganism and microbial starter. Lactobacillus plantarum has been identified from a variety of traditional foods and is characterized by systemic and molecular taxonomic purposes and produces enzymes such as -amylase, esterase, and lipase [1]. Lipase is an enzyme whose use is very broad and is found in animals, plants, fungi and bacteria. Lipases catalyze the hydrolysis of long-chain acyl glycerol at the aqueous oil interface [2]. Lipase has substrate specificity and variety different enzymatic properties, like broad source, short cycle, wide pH range, and a wide temperature range. Microbes that can produce lipases have important role than animals and plants. Lipase used in enzymatic theoretical research and practical applications for example in the process hydrolysis and esterification. Lipases from microorganisms for industrial are more attractive because lipases can be produced in large quantities and can be used as a catalyst and easily manipulated genetically. Lipases produced by microbes are heat-stable and play a role in the process of spoilage of dairy products. Factors to be considered when using microbes to produce lipases are microbial growth, the conditions of lipase production in the corresponding, test the lipase activity of the enzymes produced. Microbial growth conditions that affect lipase synthesis and production include carbon sources. Carbon sources can come from olive oil or other vegetable oils [2]. Vegetable oil has been used as an inducer for lipase production by adding vegetable oil to the media. Vegetable oils are used, such as peanut oil, sunflower oil, olive oil, palm oil and coconut oil [3]. Bacteria, yeast, and mould are known to secrete lipases to break down lipids. C. rugosa produces lipases that function to reduce serum cholesterol levels. S. marcescens has been used for the synthesis of lovastatin, a drug in which lipases are widely used for asymmetric hydrolysis of 3-phenylglycidic acid esters that are very important in the synthesis of diltiazem hydrochloride [4, 5]. MATERIALS AND METHODS Materials Lactobacillus plantarum from Microbiology and Bioprocess Technology Laboratory, Institut Teknologi Bandung, Nutrient Agar (Merck), Nutrient Broth (Merck), Aquadest sterile, 95% alcohol (Smart Lab), methyl red (Merck), palm oil (Tropical), corn oil (KCO) tween 80 (Merck), NaOH (Merck), phenolphtalein indicator (Merck), phosphate buffer pH 7 (Merck), acetone (JT baker), ethanol (Smart Lab). Methods Preparation of starter The bacterial culture of Lactobacillus plantarum made turbidity of the suspension which was arranged into a population of±1 million colonies/ml in medium nutrient agar. Then made a series that is equivalent to the turbidity of the suspension, the absorbance value of 0.8 to 1.2. Subsequently incubated at 37 ᵒC for 24-48 h. This suspension is made as a fermentation starter [6]. Making a bacterial growth curve Nutrient Broth medium was put into the 500 ml Erlenmeyer as much as 225 ml, adding a suspension of bacteria that had been made as much as 25 ml (10% v/v). Then incubated in a shaker incubator with a speed of 150 rpm for 48 h at a temperature of 35-37◦C.Sam pling was carried out every 1 hour for 48 h, then absorption measurements were carried out using UV-Vis spectrophotometry at a wavelength of 660 nm to see the turbidity level [6]. Production of bacterial metabolites Lactobacillus plantarum bacterial suspension was inoculated as much as 25 ml (10% v/v) into a 500 ml Erlenmeyer glass containing 225 ml of Nutrient Broth medium with palm oil substrate (with 1%, 3% and 5% substrate concentration variation)). Then incubated at 37 ᵒC using a shaker incubator with a speed of 150 rpm for 48 h. Sampling is done at a certain time, namely at the optimum time of secondary metabolite production based on the growth curve (until the end of the stationary phase). The fermentation results are centrifuged at 4000 rpm for 30 min to obtain a supernatant, a metabolite of the bacterium Lactobacillus plantarum [7]. Qualitative test of lipase enzyme activity Identification was carried out by inoculating bacterial metabolite fractions into 20 ml Nutrient Agar media containing 10% (2 ml w/v) International Journal of Applied Pharmaceutics ISSN- 0975-7058 Vol 13, Special Issue 3, 2021