Investigating the L-asparaginase Production in the Yeast Yarrowia ‎Lipolytica

Introduction: L-Asparaginase catalyzes the hydrolysis of L-asparagine to L-aspartic acid and ammonia. This enzyme is used for the treatment of patients with acute lymphoblastic leukemia, melanosarcoma and lymphosarcoma. It is also used in the production of acrylamide-free foods. Considering the important applications of this enzyme, the aim of this study was to introduce a new microbial source for the production of eukaryotic L-asparaginase. Materials and Methods: The quality and quantity production of L-asparaginase was investigated in four strains of yeast Yarrowia lipolytica. The quality assessment of L-asparaginase production was done on asparagine minimal agar. Quantitative assay of the enzyme was done in Czapex Dox’s medium by spectrophotometric method using Nessler’s reagent. Results: All of four strains show that they are able to produce asparaginase making a purple halo around the colony. Y. lipolytica DSM3286, DSM70562, DSM3218 and CBS6303 were produced 17/14, 11, 6/98 and 5/61 U/mL of asparaginase in Czapex Dox’s medium after 24 h, respectively. Discussion and Conclusion: Y. lipolytica DSM3286 was selected as the best asparaginase producer strain with the largest halo and the highest asparaginase production. The Y. lipolytica could be used as a potential source of L-asparaginase production

Effect of Plant Oils upon Lipase and Citric Acid Production in Yarrowia lipolytica Yeast

The nonconventional yeast Yarrowia lipolytica degrades very efficiently hydrophobic substrates to produce organic acids, single-cell oil, lipases, and so forth. The aim of this study was to investigate the biochemical behavior and simultaneous production of valuable metabolites such as lipase, citric acid (CA), and single-cell protein (SCP) by Yarrowia lipolytica DSM 3286 grown on various plant oils as sole carbon source. Among tested plant oils, olive oil proved to be the best medium for lipase and CA production. The Y. lipolytica DSM 3286 produced 34.6 ± 0.1 U/mL of lipase and also CA and SCP as by-product on olive oil medium supplemented with yeast extract. Urea, as organic nitrogen, was the best nitrogen source for CA production. The results of this study suggest that the two biotechnologically valuable products, lipase and CA, could be produced simultaneously by this strain using renewable low-cost substrates such as plant oils in one procedure

Optimization of recombinant laccase production by Yarrowia lipolytica in a medium containing glucose as carbon source with Taguchi method

Introduction:Laccases (EC 1.10.3.2; benzenediol: oxygen oxidoreductase) are copper-containing oxidases that use molecular oxygen to oxidize various aromatic and non-aromatic compounds. Laccase is applied in delignification of lignocellulosic compounds for production of bioethanol, bioremediation of industrial wastewaters especially textile, food industries, and making biosensors. Materials and methods: The Taguchi experimental design method was used for optimization of laccase production in recombinant strain Yarrowia lipolytica YL4. A L-16 Taguchi orthogonal array was used to optimize the carbon and nitrogen sources along with vitamin in four levels. Results: The results showed that glucose, ammonium chloride, yeast extract and thiamine have significant effects on the production of laccase, respectively. The laccase activity reached to 1.52 U/mL after optimization of medium which is 7.6-fold higher than un-optimized medium. Discussion and conclusion: According to the analysis of results, the Taguchi experimental design method is a successful approach to increase laccase and recombinant proteins production in Y. lipolytica

Effect of olive oil with different purity grades on Yarrowia lipolytica lipase production

 Introduction: Lipases are a class of hydrolases which catalyze the hydrolysis of triglycerides to glycerol and free fatty acids. Lipases have become an integral part of the modern food, detergents, cosmetics and pharmaceutical industries. The aim of this study was investigation of the effect of olive oil with different commercial purity as a substrate on lipase production by Yarrowia lipolytica. Materials and methods: One percentage of olive oil with three different commercial grades Extra-virgin, Virgin and Ordinary was used as cheap substrate to produce lipase by wild-type strain Y. lipolytica DSM3286 and mutant strain Y. lipolytica FDY139. Results: Maximum lipase activity was observed 72 hours after inoculation. The wild-type strain Y. lipolytica DSM3286 produced lipase with 56 U/ml activity on Virgin olive oil, but mutant strain Y. lipolytica FDY1390 produced lipase with 300 U/ml activity on Ordinary olive oil. Discussion and conclusion: The Y. lipolytica FDY1390 produced lipase about 5.4-fold higher than the wild-type strain on Ordinary olive oil. Whereas other expensive type of olive oil, Ordinary olive oil is a cheap oil which could be used to lipase production and optimization

Optimization of an Industrial Medium from Molasses for Bioethanol Production Using the Taguchi Statistical Experimental-Design Method

The production of bioethanol as a clean liquid fuel in a cost-effective way is highly desired by global energetics. Sugar beet molasses is a renewable and cheap substrate for the production of biotechnological products. Therefore, the aim of the current study was the optimization of an industrial medium from molasses for bioethanol production using the Taguchi statistical experimental-design method. First, the growth rate of yeast cells and the amount of ethanol produced by the Saccharomyces cerevisiae strain sahand 101 were investigated in aerobic and aerobic⁻anaerobic conditions. The yeast strain produced 8% (v/v) bioethanol in a medium containing molasses with 18% Brix in aerobic⁻anaerobic conditions. The main factors of the medium, including molasses, ammonium sulfate, urea, and pH, were optimized for the increase of bioethanol production by the Taguchi method. Bioethanol production reached 10% (v/v) after optimization of the medium in flask culture. The yeast strain produced 11% (v/v) bioethanol in the bioreactor culture containing the optimized medium, which is an acceptable amount of bioethanol produced from molasses at the industrial scale. The results showed that the Taguchi method is an effective method for the design of experiments aiming to optimize the medium for bioethanol production by reducing the number of experiments and time