ABSTRACT

INTRODUCTION
Lipases have come into prominance because of new and novel applications in oleo-chemistry, detergent formulation, organic synthesis and nutrition. Lipases belong to the class of serine hydrolases and therefore do not require any cofactor The natural substrates of lipases are triacylglycerols, which have very low solubility in water. Under natural conditions lipases catalyse thte hydrolysis of ester bonds at the interface between an insoluble substrate phase and the aqueous phase where the enzyme remains dissolved (Figure1). Under certain experimental conditions such as in the presence of traces of water, they are capable of reversing the reaction. The reverse recation leads to esterification and formation of glycerides from fatty acids and glycerol (fig 2). The occurence of lipase catalysed reactions at an interface between the substrate and the aqueous phase generates scientic challenges in the assay and kinetic analaysis in the reaction .

The usual industrial lipases are special classes of esterase enzymes that acts on fats and oils, and hydrolyse them into the subbstituted glycerides and fatty acids, and hydrolyse them in steps into the substituted glycerides and fatty acids, and finally on total hydrolysis into glycerol and fatty acids. BACTERIAL LIPASESgenerally bacterial lipases are glycoproteins but some extracellular bacterial lipases are lipoproteins. The production of extracellular lipases from bacteria is often dependent on nitrogen and carbon sources, inorganic salts, presence of lipids, temperature and availability of oxygen. Different genera of bacteria including streptomyces spp. produce lipase but the following genera have been well exploited for lipase production: Achromobacter spp., alcaligenes spp., arthrobacter spp., Pseudomonas spp. and Chromobacterium spp.

FUNGAL LIPASESfungi has been valued as an importantant source of lipase due to the foollowing properties : thermal stability, pH stability, substrate specificity and activity in organic solvents.Fungal lipases have benefits over bacterial ones due to the fact that present day benefits over bacterial ones due to the fact that present day technology favours the use of batch fermentation and low cost extraction methods. The chief producers of commercial lipases are aspergillus niger, A.terreus, A. carneus, Candida cylindracea, Humicola languginosa, Mucor miehei, rhizopus arrhizus, R. delemar.

Production of Extracellular Lipase from Aspergillus niger by Solid-State Fermentation

Lipases occur widely in bacteria, yeasts and fungi. Fungi are broadly recognized as one of the best lipase sources and are used widely in the

food industry. Aspergillus niger is among the most well known lipase producers. Studies on the production of extracellular lipases with A. niger have shown variations among different strains. However, the requirement for lipid carbon source remains essential for enzyme production.

SOLID STATE FERMENTATION

Solid State Fermentation (S.S.F) is a bio molecule manufacturing process used in the food, pharmaceutical, cosmetic, fuel and textile industries. These bio molecules are mostly metabolites generated by micro-organisms grown on a solid support selected for this purpose. This technology for the culture of microorganisms is an alternative to liquid or submerged fermentation, used predominantly for industrial purposes on a global scale. SSF has many advantages over submerged fermentation (SmF), including an economical use of space that is required for fermentation, simplification of the fermentation media, superior yields and no requirement for complex machinery. However, SSF has some limitations such as a poor pool of microorganisms capable of growth under restricted conditions and the controlling and monitoring of parameters such as temperature, pH, humidity and air flow. SUBSTRATE During the SmF the mineral growth medium (MGM) contained (in g/L): NaH2PO4 12, KH2PO4 2, MgSO47 H2O 0.3 and CaCl2 0.25. Ammonium sulphate at 1 % and olive oil at 2 % were used as nitrogen and carbon sources, respectively. It was also assayed with different combinations of olive oil and glucose following the feed scheme represented in Table 1.

Combination 6 was the most suitable carbon source for the Aspergillus niger lipase production.

Lipase production in SSF A mass of 5 g of wheat bran was taken in 250-mL Erlenmeyer flasks and moistened with 5 mL of sterilized salt solution of MGM (121 C for 30 min). After cooling, the flasks were inoculated with a spore suspension containing 107 spores/mL from a 7-day-old culture grown on MGM and incubated at 30 C for 7 days. After one week, 100 mL of distilled water were added to each flask and the mixture was shaken for 30 min

at room temperature to facilitate the extraction of enzyme from fermented wheat bran. At the end of the extraction, the suspension was squeezed through a double- -layered muslin cloth and it was centrifuged at 12 000 rpm for 5 min. The clear supernatant obtained was used as the extracellular enzyme.

BIOSYNTHETIC PATHWAY

IMPORTANT REGULATORY STEPS:

Effect of temperature

Effect of pH

CONCLUSION A. niger was the best producer of extracellular lipase and its synthesis is induced by the presence of a lipid source. Moderate enzyme activity (9.14 IU/g dss) was obtained in SSF using wheat bran, are amongst the highest extracellular lipase activities reported in the literature concerning fungal sources. This lipase has some properties in common with lipases from other A. niger strains. This enzyme is also stable over a broad pH range, 4 to 7, for a period of 24 hours at 30 C. Due to the fact that these microorganisms are generally recognized as safe (GRAS) for food, brewing and pharmaceutical applications, more research is necessary

to optimize the fermentative process in order to obtain higher lipase production through this strain. REFRENCES

Production of Extracellular Lipase from Aspergillus niger by Solid-State Fermentation

Gwen Falony1, Janny Coca Armas1*, Julio C. Dustet Mendoza1 and Jos L. Martnez Hernndez2**

http://www.wikipathways.org/index.php/Pathway:WP71 http://nopr.niscair.res.in/bitstream/123456789/5772/1/IJBT%204(4)%20437-445.pdf