A study of the enzymatic profile of soil isolates of Nocardia asteroides

In this study, using the API-ZYM system, we have reported the enzyme profile of 42 soil strains and 2 clinical strains of Nocardia asteroides isolated locally. Of the 19 enzymes tested, only 7 were demonstrable in over 90% of the soil isolates. These included alkaline phosphatase, esterase lipase, leucine arylamidase, acid phosphatase, phosphohydrolase, α-glucosidase and β-glucosidase. In addition, β-galactosidase activity was demonstrated in all the strains by the O-nitrophenyl-β-D-galactopyranoside (ONPG) test. The enzymes which were not demonstrable in >95% of the strains included valine arylamidase, cystine arylamidase, trypsin, chymotrypsin, α-galactosidase, β-glucoronidase, N-acetyl-β-glucosaminidase, α-mannosidase and α-fucosidase. With the exception of valine arylamidase, which was lacking in all but one isolate, the enzyme profiles of the soil isolates were comparable with the clinical isolates of N. asteroides reported in previous studies. The reasons for this difference in the two sets of isolates is not clear. The study reinforces the view that specific differences in the enzymatic profiles of Nocardia species could be used for their rapid identification. However, more extensive studies are needed to establish the reproducibility of this method. To the best of our knowledge, this is the first study of the enzymatic profile of soil isolates of N. asteroides originating from a single geographic region. This revised version was published online in June 2006 with corrections to the Cover Date.     

Extracellular enzymatic activity ofMicrosporum canis isolates

The enzymatic activity of 70 feline and canineMicrosporum canis isolates was determined by the Api-Zym^® test. The liquid phase of cultures, inoculated into Tryptic Soy Broth, was used to examine 19 enzymes. Considerable differences were observed among the extracellular enzymatic patterns. All the isolates produced alkaline phosphatase and beta-glucosidase, while lipase (C14), trypsin, chymotrypsin, beta-glucuronidase, and alpha-fucosidase activity was never revealed. Esterase (C4) activity was present in 57 samples (81%), esterase lipase (C8) in 31 (44%), leucine arylamidase in 35 (50%), valine arylamidase and cystine arylamidase in 7 (10%), acid phosphatase in 64 (91%), naphthol-AS-BI-phosphohydrolase in 60 (86%), alpha-galactosidase in 5 (7%), beta-galactosidase in 6 (8%), alpha-glucosidase in 25 (36%), N-acetyl-beta-glucosaminidase in 41 (58%), and alpha-mannosidase in 51 (73%). The beta-galactosidase activity ofM. canis has not been reported previously. Remarkable variations of intensity for each enzymatic activity were also detected. It is believed that these results could provide basic data for further investigations on the pathogenic role of enzymes secreted byM. canis.     

Digestion in the soil predatory mite Pergamasus longicornis (Berlese) (Acari: Mesostigmata: Parasitidae)--detectable hydrolases

Nineteen hydrolytic enzymes were detected in individual adult Pergamasus longicornis (Berlese) mites--amylase, hide protease, alkali phosphatase, esterase (C4), esterase lipase (C8), lipase (C14), leucine arylamidase, valine arylamidase, cystine arylamidase, acid phosphatase, phosphoamidase, alpha-galactosidase, beta-galactosidase, beta-glucuronidase, alpha-glucosidase, beta-glucosidase, N-acetyl-beta-glucosaminidase, alpha-mannosidase, and alpha-fucosidase. All but the phosphatases were detected for the first time. Tryptic and chymotryptic activity were consistently not demonstrable. Comparisons are made with saprophagous mites. No clear enzymic specialization for predation was found.     

Enzyme activities ofBorrelia burgdorferi sensu lato

Activities of 19 enzymes were tested by the API ZYM system in 13 strains ofBorrelia burgdorferi sensu lato (B. burgdorferi sensu stricto,B. afzelii, B. garinii, B. lusitaniae, B. valaisiana) grown in liquid BSK-H medium supplemented with rabbit serum. All strains produced acid phosphatase, esterase (C4), esterase-lipase (C8), leucine arylamidase and naphthol-AS-BI-phosphohydrolase. Nine strains also produced alkaline phosphatase, and three strains produced α-glucosidase. The API ZYM system probably cannot be used for differentiation betweenB. burgdorferi sensu lato genomospecies.     

Extracellular enzymatic activities in Cryptococcus neoformans strains isolated from AIDS patients in different countries

Three hundred and ten Cryptococcus neoformans strains isolated from AIDS patients in five different countries (151 from Brazil, 23 from Italy, 28 from Spain, 104 from Thailand and four from Turkey) were tested by the API-ZYM kit to detect their extracellular enzymatic activity. The enzymes esterase (C4) (no 3), esterase lipase (C8) (no 4), leucine arylamidase (no 6) and acid phosphatase (no 11) were commonly positive in most of the strains (more than 95%). These enzymes could be considered a useful tool not only for C. neoformans identification, but in particular for their possible relationship to new C. neoformans virulence factors and also for epidemiological research. Interestingly, it is also the high positive percentage of alpha-glucosidase and beta-glucosidase detected in all isolates. The serotype A was the most predominant serotype in all countries, except for Italy where the serotype D was predominant. Further studies are needed to draw a clear correlation between the API-ZYM profile and serotype.     

Candida species in tchapalo and bangui, two traditional alcoholic beverages from Côte d'Ivoire

The increase of infections due to non-Candida albicans species made it very necessary to conduct adequate characterization to be able to identify the species of Candida isolated from traditional fermented foods. In this study, based on their hue on Candida Chromogenic Agar medium, a total of 136 yeast strains were isolated from tchapalo and bangui. Molecular identification based on PCR-RFLP of internal transcribed spacers of rDNA (ITS) and sequencing of the ITS and the D1/D2 regions allowed us to assign these isolates to seven species: Candida tropicalis, Candida inconspicua, Candida rugosa, Saccharomyces cerevisiae, Kluyveromyces marxianus, Hanseniaspora guilliermondii, Trichosporon asahii. With the respect to each beverage, six species were found among with four species are regarded as opportunistic pathogens. From these, C. tropicalis, C. inconspicua and K. marxianus were the most commonly encountered. The enzyme activities of the potential pathogens assessed using API ZYM system showed that almost strains had esterase, esterase lipase, valine and cystine arylamidase, alpha chymotrypsin, alkaline phosphatase and naphthol phosphohydrolase activities. The activity of α-glucosidase was found only in C. tropicalis and C. inconspicua strains isolated from tchapalo while β-glucosidase activity was found in all strains from tchapalo and only in C. inconspicua isolated from bangui.     

Activity of hydrolytic enzymes of Candida albicans strains isolated from patients with periodontal and membrane mucosae of oral cavity diseases

Fungi are elements of the ontocenosis of the oral cavity and causal factors of inflammatory lesions in its mucous membrane. The objective of the study was to find differences in the activity of hydrolytic enzymes of Candida albicans isolated from patients with diseases of the periodontium and mucous membrane of the oral cavity. Of 235 patients examined, 31 were diagnosed with gingivitis, 38 with glossitis, 28 with leucoplakia, 37 with adult periodontitis, 25 with juvenile periodontitis, 36 with stomatitis prothetica and 40 with stomatitis atrophica. In 196 patients (83.4 ± 2.4%), fungi belonging to Candida species were detected. In the evaluation of Candida albicans strains (146) properties, bioMerieux API ZYM tests containing substrates for the detection of 19 hydrolases were used. All the investigated strains were characterized by the activity of 14 enzymes, i.e. phosphatase alcaline, esterase (C4), esterase lipase (C8), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, α galactosidase, β galactosidase, α glucosidase, β glucosidase, N-acetyl-β-glucosaminidase, α mannosidase and α fucosidase. Strains isolated from the oral cavity of patients with diseases of periodontium and mucous membrane are characterised by the highest phosphatase acid activity. The greatest enzymatic activity is characteristic of Candida albicans isolated from patients with stomatitis atrophica or stomatitis prothetica, and the lowest in strains from gingivitis or juvenile periodontitis cases. Differences in the activity of hydrolases are statistically significant (p<0.01) for: esterase (C4), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, β glucosidase, N-acetyl-β-glucosaminidase, of fungi isolated from patients with particular clinical diagnoses. This revised version was published online in June 2006 with corrections to the Cover Date.     

Characterization of Common Carbohydrate Antigenic Determinants on Soya Bean Cell-Wall Enzymes

Three soya-bean (Glycine max) cell-wall enzymes (ß-glucosidase,pectin methyl esterase and phosphatase) have been found to beglycoproteins. The polyclonal antibodies raised against pectinmethyl esterase and ß-glucosidase lacked specificity,cross-reacted highly with native enzymes and also both reactedwith pure soya-bean phosphatase, horseradish peroxidase andhoneybee venom phospholipase A2. They did not react with eithernon-glycosylated bacterial phosphatase or deglycosylated cell-wallenzymes. The two antisera contained both non-specific anti-glycanantibodies and specific anti-polypeptide antibodies that werequantified. Antiglycan antibodies specific to 1–3 fucoseand ß1–2 xylose were detected in both antiseraand were separated and quantified. The occurrence of terminalfucose (and mannose) was confirmed with specific lectins. Theseresults indicate that most of the common glycan epitopes probablycorrespond to the asparagine-linked complex glycan previouslydetected in several glycoproteins of plants as well as in thoseof molluscs and insects. (Received March 10, 1993; Accepted November 5, 1993)     

Secretory Hydrolases of Entamoeba histolytica1

ABSTRACT Cells of Entamoeba histolytica grown over a period of four days contained NADP⁺-dependent alcohol dehydrogenase exclusively inside the cells. No activity of this enzyme could be found in the growth medium after harvesting the cells. Under the same conditions, acid phosphatase, β-N-acetylglucosaminidase, esterase, α-glucosidase, and different amylases of the parasite were found both inside the cells and in the medium. The activities present in the cell homogenate and in the medium before and after growth of the amoebas were partially separated by gel filtration on Sephadex G150 and G75, respectively. The comparison of the elution diagrams revealed that NADP⁺-dependent alcohol dehydrogenase, acid phosphatase, esterase, and amylases occurred as multiple forms inside the cells. These activities, as well as β-N-acetylglucosaminidase and α-glucosidase, were released into the extracellular environment to a different degree. The enzymes originating from the parasite were identified and distinguished from those of the ingredients of the growth medium according to their molecular mass and pH optimum. Furthermore, the amoebic origin of the secreted enzymes was shown on the basis of their inhibition by antibodies prepared against the supernatant fraction of the homogenate.     

Detection,isolation and characterization of Fusobacterium gastrosuis sp. nov. colonizing the stomach of pigs

Nine strains of a novel Fusobacterium sp. were isolated from the stomach of 6–8 months old and adult pigs. The isolates were obligately anaerobic, although they endured 2 h exposure to air. Phylogenetic analysis based on 16S rRNA and gyrase B genes demonstrated that the isolates showed high sequence similarity with Fusobacterium mortiferum, Fusobacterium ulcerans, Fusobacterium varium, Fusobacterium russii and Fusobacterium necrogenes, but formed a distinct lineage in the genus Fusobacterium. Comparative analysis of the genome of the type strain of this novel Fusobacterium sp. confirmed that it is different from other recognized Fusobacterium spp. DNA-DNA hybridization, fingerprinting and genomic %GC determination further supported the conclusion that the isolates belong to a new, distinct species. The isolates were also distinguishable from these and other Fusobacterium spp. by phenotypical characterization. The strains produced indole and exhibited proline arylamidase and glutamic acid decarboxylase activity. They did not hydrolyse esculin, did not exhibit pyroglutamic acid arylamidase, valine arylamidase, α-galactosidase, β-galactosidase, β-galactosidase-6-phosphate or α-glucosidase activity nor produced acid from cellobiose, glucose, lactose, mannitol, mannose, maltose, raffinose, saccharose, salicin or trehalose. The major fatty acids were C16:0 and C18:1ω9c. The name Fusobacterium gastrosuis sp. nov. is proposed for the novel isolates with the type strain CDW1(T) (=DSM 101753(T) = LMG 29236(T)). We also demonstrated that Clostridium rectum and mortiferum Fusobacterium represent the same species, with nomenclatural priority for the latter.     

Contribution to the study of the enzymatic profiles of yeast organisms with medical interest

The enzymatic profiles of several yeastlike organisms were studied using 19 substrates included in the API ZYM system. The isolates evaluated were: 186 Candida albicans, 19 C. stellatoidea, 4 C. tropicalis, 2 C. parapsilosis, 2 C. pseudotropicalis, 1 C. guilliermondii, 3 C. krusei, 11 Torulopsis (Candida) glabrata, 1 Cryptococcus neoformans, 2 Saccharomyces carlsbergensis, 1 Rhodotorula rubra, and 1 R. mucilaginosa. Esterase lipase (C8), leucine arylamidase, acid phosphatase, and phosphoamidase were detected in all of the isolates while trypsin and alpha-galactosidase were not found in any of the isolates using this system. The other enzymes were produced to a variable degree. The different enzymatic profiles might prove useful in the rapid differential diagnosis of genera and species of these yeastlike organisms. To this end, more extensive studies using more isolates of each species will be required, and enzymatic activity should be verified with other techniques and substrates.     

Molecular characterization of European and Egyptian isolates of Sclerotium cepivorum,the incitant of onion white rot

Abstract

The tested European and Egyptian isolates of Sclerotium cepivorum were able to infect Giza 6 onion cultivar causing white rot disease with a different degrees of disease severity (ranging from sever to weak). The pattern of esterase isozymes produced by the tested isolates of the pathogen showed two main bands (arrows) which were different in density. Such differences in density of bands were present in every run and therefore appear to be indicators for differences among the tested isolates. Analysis of the protein pattern of the tested isolates of the pathogen indicated that the tested isolates had major proteins of a molecular weight of 52, 36, 23 and 16 kDa. Variation between isolates was detected by presence of bands of low molecular weight. Isolate Nos. 1, 4, 5, 7, 8, 9, 10 and 13 had a band at 17 kDa, whereas isolate Nos. 2, 3, 6, 11, 12, 14, and 15 had a band at 20 kDa. Using RAPD analysis to evaluate the genetic diversity of the tested isolates indicated that the tested field population of the pathogen was genetically heterogeneous but shared a number of common bands with molecular weights ranging from 650 to 2500 bp. Based on the DNA banding pattern the tested isolates can be assigned to seven genetically different groups. All tested isolates produced a band at 2500 bp except isolate No. 7. No correlation was exibited between patterns esterase isozmes, protein and DNA patterns of S. cepivorum isolates and their virulence or geographical origin.     

Enzyme activities of fungi associated with Picea abies needles

Decomposition of Picea abies needles and production of extracellular enzymes involved in decomposition of lignin, cellulose, hemicelluloses and other organic compounds were studied in fungal strains of interior needle colonizers isolated from needles in different stages of decomposition (attached to trees, and early and late decomposition stages in the litter horizon). In total, 12 strains of ascomycetes (members of Helotiales, Hypocreales, Dothideales, Diaporthales and Eurotiales) and four basidiomycetes (Polyporales, Agaricales and Russulales) were tested. Significant decomposition of needles was recorded for all fungal isolates. All isolates produced cellobiohydrolase, β-glucosidase, β-xylosidase, N-acetylglucosaminidase, α-glucosidase, phosphatase and arylsulfatase and most fungi also produced endocellulase, endoxylanase and laccase in needle litter. In addition, other hemicellulases were produced by all strains. Mn-peroxidase was only produced by two basidiomycetes. Although enzyme activities varied, fungi associated with needles on fallen trees exhibited enzyme production comparable with later litter colonizers, and there was no significant difference in enzyme production between ascomycete and basidiomycete strains.     

Subcellular fractionation of midgut cells of the sunn pest Eurygaster integriceps (Hemiptera: Scutelleridae): Enzyme markers of microvillar and perimicrovillar membranes

The subcellular distributions of six digestive and non-digestive enzymes (α-glucosidase, β-glucosidase, alkaline phosphatase, acid phosphatase, aminopeptidase and lactate dehydrogenase) of Eurygaster integriceps have been studied. The subcellular distributions of acid phosphatase and α-glucosidase are similar and the gradient ultracentrifugation profiles of these two enzymes overlap. Two partially membrane-bound enzymes, alkaline phosphatase and β-glucosidase have similar distributions in differential centrifugation fractions, which are different from that of α-glucosidase. Sucrose gradient ultracentrifugation of membranes from luminal contents showed that β-glucosidase carrying membranes are heavier. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the profile of proteins extracted from β-glucosidase carrying membranes is different from that of α-glucosidase carrying membranes. We conclude that β-glucosidase and aminopeptidase are markers of microvillar membrane (MM) and perimicrovillar space, respectively, while α-glucosidase and acid phosphatase are perimicrovillar markers. In E. integriceps V1 luminal content is a rich source of PMM and MM and that is used to resolve these membranes.     

Acid and Neutral Hydrolases in Trypanosoma cruzi. Characterization and Assay*

Twelve acid hydrolases, 4 near-neutral hydrolases and alkaline phosphatase were demonstrated in 0.34 M sucrose homogenates of Trypanosoma cruzi strain Y: p-nitrophenylphosphatase and α-naphthylphosphatase, with optimum pH at ? 6.0; α-galactosidase, β-galactosidase, β-glucosidase, N-acetyl-β-glucosaminidase, cathepsin A and peptidase I and III, with optimum pH between 5.0 and 6.0: and arylsulfatase cathepsin D, α-arabinase and α-mannosidase with optimum pH at ? 4.0 α-Glucosidase, gluccse-6-phosphatase and peptidase II had optimum pH at ? 7.0. β-Glycerophcsphatase had a broad pH-activity curve from 4.0 to 7.4, with maximum activity at pH 7.0. The main kinetic characteristics of these enzymes and their quantitative assay methods were studied. No activity was detected for α-fucosidase, β-xylosidase, β-glucuronidase, elaidate esterase. acid lipase, and alkaline phospho-diesterase.     

Main airborne Ascomycota spores: characterization by culture, spore morphology, ribosomal DNA sequences and enzymatic analysis

The aim of this work was to identify the main allergy-related Ascomycetes fungal spores present in the atmosphere of Porto, using different and complementary techniques. The atmospheric sampling, performed in the atmosphere of Porto (Portugal) from August 2006 to July 2008, indicated Cladosporium, Penicillium, Aspergillus and Alternaria as the main fungal spore taxa. Alternaria and Cladosporium peaks were registered during summer. Aspergillus and Penicillium highest values were registered from late winter to early spring. Additionally, the Andersen sampler allowed the culture and isolation of the collected viable spores subsequently used for different identification approaches. The internal-transcribed spacer region of the nuclear ribosomal repeat unit sequences of airborne Ascomycetes fungi isolates revealed 11 taxonomically related fungal species. Among the identified taxa, Penicillum and Aspergillus presented the highest diversity, while only one species of Cladosporium and Alternaria, respectively, were identified. All selected fungal spore taxa possessed phosphatase, esterase, leucine arylamidase and β-glucosidase enzymatic activity, while none had lipase, cystine arylamidase, trypsin or β-glucuronidase activity. The association between the spore cell wall morphology, DNA-based techniques and enzymatic activity approaches allowed a more reliable identification procedure of the airborne Ascomycota fungal spores.     

Extracellular hydrolase profiles of fungi isolated from koala faeces invite biotechnological interest

The extracellular enzymes of seven fungal strains isolated from koala faeces have been comprehensively characterised for the first time, revealing potential for biotechnological applications. The fungal isolates were grown in a hydrolase-inducing liquid medium and the supernatants were analysed using enzyme assays and zymogram gels. Temperature and pH profiles were established for xylanase (EC 3.2.1.8 endo-1,4-β-xylanase), mannanase (EC 3.2.1.78 mannan endo-1,4-β-mannosidase), endoglucanase (EC 3.2.1.4 cellulase), β-glucosidase (EC 3.2.1.21 β-glucosidase), amylase (EC 3.2.1.1 α-amylase), lipase (EC 3.1.1.3 triacylglycerol lipase) and protease (EC 3.4 peptidase) activities. Comparisons were made to the high-secreting hypercellulolytic mutant strain Trichoderma reesei RUT-C30 and the wild-type T. reesei QM6a. The isolates from koala faeces Gelasinospora cratophora A10 and Trichoderma atroviride A2 were good secretors of total protein and heat-tolerant enzymes. Doratomyces stemonitis C8 secreted hemicellulase(s), endoglucanase(s) and β-glucosidase(s) with neutral to alkaline pH optimums. A cold-tolerant lipase was secreted by Mariannaea camptospora A11. The characteristics displayed by the enzymes are highly sought after for industrial processes such as the manufacture of paper, detergents and food products. Furthermore, the enzymes were produced at good starting levels that could be increased further by strain improvement programs.     

Histochemistry of the development of the digestive system of Siberian sturgeon during early ontogeny

At hatching, the yolk-sac matrix of Siberian sturgeon Acipenser baeri contained neutral glycoconjugates, glycogen, proteins rich in arginine, lysine, tyrosine, cysteine and cystine, glycoproteins containing mannose (Man) and/or glucose (Glc), N -acetyl-D-galactosamine (GalNAc), L-fucose (Fuc), sialic acid and/or N -acetyl-D-glucosamine (GlcNAc) residues, as well as neutral and acidic lipids. Buccopharyngeal and anterior oesophageal goblet cellls produced a combination of neutral and acid sialoglycoproteins, while those from the posterior oesophagus secreted only neutral glycoproteins; both types of secretions contained tryptophan and -S-S- groups and were unreactive to lectin techniques. Most intestinal goblet cells secreted mainly carboxylated and sulphated sialoglycoproteins with some rests of neutral glycoconjugates, while few of them produced only acid or neutral glycoproteins. Intestinal glycoproteins were rich in GalNAc, GlcNAc and sialic acid residues. Close relationships between digestive enzymes and morphological development of digestive organs were observed. Histochemistry of enzymes revealed that just after hatching, alkaline and acid phosphatase, ATP -ase and non-specific esterase activities were detected in the yolk sac. From the onset of exogenous feeding to the juvenile stage (30 days post-hatch), an enhancement of enzymatic activities was observed, as alkaline and acid phosphatase, ATP -ase, aminopeptidase M and nonspecific esterase sharply increased. However, lipase activity decreased in the liver and brush border of enterocytes by 13–14 days post-hatch. Two types of lipase were detected in the alimentary canal, a non-pancreatic lipase that was secreted in the cardiac stomach by gastric glands, and a pancreatic lipase, which activity was mainly detected in the brush border of the intestinal epithelium.     

Probiotic Characteristics of <Emphasis Type="Italic">Lactobacillus curvatus</Emphasis> DN317, a Strain Isolated from Chicken Ceca

The probiotic characteristics of Lactobacillus curvatus DN317, a strain isolated from chicken ceca, were evaluated. This strain was selected for study from the isolated Lactobacillus strains because it has specific anti-microbial activity against Campylobacter jejuni ATCC 33560, Camp. jejuni NCTC 11168, Listeria monocytogenes ATCC 7644, and Bacillus subtilis ATCC 8633. Lact. curvatus DN317 showed an auto-aggregation percentage of 72% and presented the highest co-aggregation with Lact. monocytogenes ATCC 7644 (68%) compared to B. subtilis ATCC 8633 (45%), Camp. jejuni ATCC 33560 (36%), and Camp. jejuni NCTC 11168 (35%). The data revealed that Lact. curvatus DN317 could survive at 0.25% bile, maintain viability at pH 2.5 for 30 min, produce biosurfactants, and adhere to Caco-2 cells. Quantification of IL-6, IL-8, IL-10, and β-defensin 2 levels shows that Lact. curvatus DN317 induces an increase in IL-8 and β-defensin 2 secretion, while the levels of IL-6 and IL-10 do not change. Lact. curvatus DN317 showed high levels of esterase and cysteine arylamidase activities (5); moderate levels of esterase lipase, β-galactosidase, and α-galactosidase activities (4, 3); and weak levels of leucine arylamidase, valine arylamidase, and acid phosphatase activity (1). Various activities were obtained of α-chymotrypsin, β-glucuronidase, β-glucosidase, and N-acetyl-β-glucosaminidase, which have been associated with intestinal diseases. Lact. curvatus DN317 lowered the cholesterol level in MRS with and without bile. Antibiotic susceptibility tests indicated that DN317 was sensitive to ampicillin, gentamicin, kanamycin, streptomycin, tetracycline, clindamycin, erythromycin, and vancomycin but was resistant to chloramphenicol and ciprofloxacin. These results suggest that Lact. curvatus DN317 could potentially function as a probiotic.     

Media Formulation using Complex Organic Nutrients for Improved Activity,Productivity, and Yield of Candida rugosa Lipase and Esterase Enzymes

Abstract

Candida rugosa is an excellent source of multiple lipase and esterase enzymes; therefore, it is of technological importance to formulate the medium that provides high activity for each enzyme. In this work, the cultivation medium comprising complex nutrients that provided the highest activity, productivity, and yield of C. rugosa enzymes individually was formulated. Time courses of the extracellular and intracellular lipase and esterase activities of C. rugosa were represented and the role of protease in the cultivation progress was discussed. Urea, soy-peptone, yeast extract, a mixture of soy-peptone and yeast extract, cheese whey, and wheat mill bran were tested for their lipolytic and esterasic activities. Urea provided considerably higher extracellular lipase activity when compared to other nitrogen sources; however, soy-peptone provided the highest extracellular esterase activity. Hazelnut, olive, sesame, soybean, and flax seed oils affected the enzyme activities to different extents related to their fatty acid compositions. Hazelnut oil and olive oil provided the highest extracellular lipase and esterase activities, respectively, whereas sesame oil produced the highest biomass. High C₁₈ and C₁₆ ester contents of vegetable oils promoted high lipase and esterase productions, respectively. A temperature of 30°C yielded the highest extracellular and intracellular lipase and esterase activities; however, 35°C produced the highest biomass.