Lipase production in continuous culture of Candida rugosa

Abstract

The yeast Candida rugosa produces multiple extracellular lipases. The production of extra-and intracellular lipases was investigated in continuous cultures using a sole or different mixtures of carbon sources. Also, the effect of different C:N ratios was tested. Lipase productivity in continuous cultures increased by 50% compared with data obtained from batch fermentations and depended on the dilution rate applied. Maximum yields relative to consumed substrate were obtained with oleic acid at low dilution rate. It was found that during nitrogen limitation, lipase activity was suppressed. All carbon source mixtures tested allowed both cell growth and lipase production, but extra-and intracellular lipase activities were affected by the combination of substrates used. Maximum extracellular lipolytic productivity was attained with lactic and oleic acid mixtures, probably due to the non-repressor effect of these carbon sources. The chemical composition of the biomass also depended on the type of substrate used and was related to the accumulation of lipidic compounds as intracellular inclusions, which were observed when oleic acid was used as the carbon source. The results obtained were compared with previous data from batch and fed-batch cultures in order to select the best process strategies for the lipase production with C rugosa. The best lipase yields were obtained in fed-batch fermentations using oleic acid.

Industrial derivative of tallow: a promising renewable substrate for microbial lipid, single-cell protein and lipase production by Yarrowia lipolytica

“…In the present study, the simultaneous increase of carbon and nitrogen concentrations notably increased lipase yield (Lip max = 2.50 IU/ml). In accordance with our findings, Montesinos et al (2003) studying the effect of nitrogen limitation upon lipase secretion by Candida rugosa ATCC found that Lip secretion was almost completely suppressed in nitrogenlimited media. Pereira-Meirelles et al (1997) reported maximum lipase concentrations around 2.5-3.0 IU/ml by a wild C. lipolytica strain cultivated on media containing olive oil and peptone, while higher quantities of total (extra-cellular and cell-bound) lipase were obtained in media enriched with both yeast extract and peptone (5.0-8.5 IU/ml) (Pereira-Meirelles et al 2000).…”

“…Results for other wild Candida species showed a lipase production of the same magnitude with the present investigation (e.g. concentrations 2-15 IU/ml for various carbon sources used) (Dalmau et al 2000;Montesinos et al 2003). In order to obtain increased lipase production by Y. lipolytica strains, combinations of genetic engineering and mutagenesis methods have been employed.…”

“…The simultaneous production of microbial lipid and lipase has rarely been reported (Szczęsna- Antczak et al 2006). A wide range of microorganisms has been reported capable of producing lipases with different enzymological properties and substrate specificities (Hadeball, 1991;Dalmau et al 2000;Sharma et al 2001;Montesinos et al 2003;Szczęsna-Antczak et al 2006). Among them, various Y. lipolytica strains have been tested, with secretion of extracellular lipase being stimulated by the presence of longchain fatty acids or triacylglycerols as carbon source and organic nitrogen (i.e.…”

Yarrowia lipolytica: A model microorganism used for the production of tailor‐made lipids

“…), while their production is also substantially enhanced by the addition of organic nitrogen (e.g., peptone, urea, etc.) to the culture medium [66,83,84,[86][87][88][89]. In the submerged cultures carried out with stearin utilized as the sole substrate by the strain ACA-DC 50109, extracellular lipase was secreted into the medium in non-negligible amounts; lipase quantities were detected in the culture medium regardless of the substrate fat or the nitrogen concentration, and secretion occurred during primary anabolic activity [40].…”

Culture Conditions and Investigation of Bioreactor Configurations for Lipase Production by Rhizopus oryzae

“…However, a considerable amount of the research carried out to date has focused on small-scale procedures, while less information is available on the particular characteristics of lipolytic enzyme biosynthesis in bioreactors and the feasibility of diverse culture modes, such as batch, fed-batch, or continuous systems [6][7][8]. Over the last few months, research concerning lipase production by yeasts and fungi in bioreactors has been employed to obtain high levels of lipase production, thus demonstrating the interest that the scale-up of this kind of biological process is triggering nowadays and then encouraging further research in this field [9][10][11][12].…”