Highest CMCase Research Articles

Cellulolytic Potential of Actinomycetes Isolated from Different Habitats

Twenty actinomycete isolates capable of utilizing lignocellulosic biomass were isolated from different samples of agricultural field soil, compost and decaying vegetation. Qualitative screening of all isolates during plate assay method using Congo red showed 17 of them producing a clear zone of hydrolysis around the colonies indicating cellulose hydrolytic potential of the isolates. Quantitative estimation of filter paper (FPase) and carboxymethylcellulase (CMCase) activities exhibited by the positive isolates showed that highest CMCase was produced by NAA2 (0.366 IU/ml), while highest FPase activity of 0.075 FPU/ml was observed in NAA14. All of the isolates showed FPase activities comparable to that shown by C. fimi (0.023 FPU/ml) taken as a standard, confirming the cellulose degradation ability of the isolates.

Isolation of cellulolytic microcosms from bagasse compost in co-digested fibrous substrates

Symbiotic cellulolytic microcosms represent a promising microbial agent for enhancing degradation of cellulosic materials in biotechnological processes. In this study, structurally stable lignocellulose-degrading microbial communities were constructed from cellulolytic seed culture from sugarcane bagasse compost using swine manure and Napier grass as co-digested carbon sources under static aerobic condition at 55 °C. The lignocellulolytic microbial consortium enriched in peptone and yeast extract-based medium (PLMC) showed higher cellulose-degrading activity compared to lignocellulolytic microbial consortium isolated in the water-based medium (WLMC). The composite microbes in both consortia were originated from the seed culture and the co-digested substrates according to denaturing gradient gel electrophoresis profile. PLMC exhibited higher CMCase, xylanase, FPase, and avicelase activities in the supernatant than those of WLMC and was capable of degrading 70 % of filter paper within 1 week. PLMC was capable of degrading substrate with higher efficiency than the control by 11.7, 9.2, 15.5, and 11.9 % of total solid, suspended solid, volatile solid, and volatile suspended solid, respectively. The work demonstrated the potential of cellulolytic microcosms enriched by this approach on enhancing conversion efficiency in biogas production from cellulosic wastes.

Optimization of Conditions and Production of Carboxy Methyl Cellulase by Bacteria Isolated from Higher Termite Soil

The current work deals with the studies of isolation and preliminary characterization of the bacteria isolated from higher termite soil. Termites play an important role in the turnover and mineralization of complex biopolymers, such as wood and other cellulose and hemicelluloses containing materials. Lignocellulose is the most predominant component of the woody and dead plant materials, as well as it is the most abundant biomass on earth, especially in terrestrial ecosystems. Thus the work focuses on using lingo cellulosic waste efficiently for production of Carboxy Methyl Cellulase (CMC). As CMCase have higher hydrolyzing capacity than the other two cellulases, cellobiase and cellobiohydralases it has wide application in industry such as bread and baking industry, laundry etc. Thus the work focuses on identification of efficient bacterial species in this work 6 bacterial species were isolated among which I4 produced higher CMCase enzyme. After the morphological, biochemical and identification test, the I4 isolate was found to be Bacillus sp.

Comparative characterization of proteins secreted by Neurospora sitophila in solid-state and submerged fermentation

Although submerged fermentation (SmF) accounts for most of current enzyme industries, it has been reported that solid-state fermentation (SSF) can produce higher enzyme yields in laboratory scale. In order to understand the reasons contributing to high enzyme production in SSF, this study compared the cellulase activities and secretomes of Neurospora sitophila cultured in SSF and SmF using steam exploded wheat straw as carbon source and enzyme inducer. The total amounts of protein and biomass (glucosamine content) in SSF were respectively 30 and 2.8 times of those inSmF. The CMCase, FPA and β-glucoside activities in SSF were 53-181 times of those in SmF. Both in SSF and SmF,N.sitophila secreted the most critical cellulases and hemicellulases known for Trichoderma reesei, although a β-xylosidasewas exclusively identified in SSF. Six endoglucanases were identified in N.sitophila secretion with the high CMCase activity. The non-enzyme proteins in SSF were involved in fungal mycelia growth and conidiation; while those in SmF were more related to glycometabolism and stress tolerance. This revealed that SSF more likely serves as a natural habitat for filamentous fungi to facilitate the enzyme secretion.

Saccharification of Sugarcane Bagasse by Enzymatic Treatment for bioethanol production

Aims: The escalating demands for traditional fossil fuels with unsecured deliverance and issues of climate change compel the researchers to develop alternative fuels like bioethanol. This study examines the prospect of biofuel production from high carbohydrate containing lignocellulosic material, e.g. sugarcane bagasse through biological means. Methodology and Results: Cellulolytic enzymes were collected from the culture filtrate of thermotolerant Trichoderma viride grown on variously pretreated sugarcane bagasse. CMCase and FPase enzyme activities were determined as a measure of suitable substrate pretreatment and opt imum condition for cellulolytic enzyme production. The highest CMCase and FPase activity was found to be 1.217 U/ml and 0.109 U/ml respectively under the production conditions of 200 rpm, pH 4.0 and 50 °C using steamed NaOH treated bagasse as substrate. SEM was carried out to compare and confirm the activity of cellulolytic enzymes on sugarcane bagasse. Saccharification of pretreated bag asse was carried out with crude enzymes together using a twofactor experimental design. Under optimized conditions the pretreated bagasse was saccharified up to 42.7 % in 24 h. The hydrolysate was concentrated by heating to suitable concentration and then used for fermentation by an indigenous isolate of Saccharomyces cerevisiae . With 50 and 80 % brix containing liquor the concentration of alcohol was 0.579 % and 1.15 % respectively. Conclusion, significance and impact of study: This is the first report in Bangladesh for the production of cellulosic ethanol using local isolates. Though the rate of alcohol production was very low, a great impetus in this field can maximize the production thereby meet the demand for fuel in future.

In vitro and in situ activity of carboxymethyl cellulase and glutamate dehydrogenase according to supplementation with different nitrogenous compounds

Two experiments were carried out to evaluate the effect of supplementation with different nitrogenous compounds on the activities of carboxymethil cellulase (CMCase) and glutamate dehydrogenase (GDH). In the first experiment, four treatments were evaluated in vitro: cellulose, cellulose with casein, cellulose with urea, and cellulose with casamino acids. After 6, 12 and 24 hours of incubation, CMCase and GDH activity, pH, and concentrations of ammonia nitrogen (AN) and microbial protein were measured. In the three incubation periods, the concentration of AN was higher when urea was used as a supplemental source of nitrogen. The activity of CMCase was higher with the addition of urea and casamino acids when compared with the control and the casein treatment. Supplementation with casamino acids provided higher GDH activity when compared with the control at 6 hours of incubation. At 12 hours of incubation, the GHD activity was also stimulated by casein. At 24 hours, there was no difference in GHD activity among treatments. In the second experiment, three rumen-fistulated bulls were used for in situ evaluation. Animals were fed Tifton hay (Cynodon sp.) ad libitum. The treatments consisted of control (no supplementation), supplementation with non-protein nitrogenous compounds (urea and ammonium sulphate, 9:1) and supplementation with protein (albumin). In treatments with nitrogenous compound supplementation, 1 g of crude protein/kg of body weight was supplied. The experiment was conducted in a 3 × 3 Latin square design. The measurements were performed at 6, 12 and 24 hours after supplementation. No difference in GDH activity was observed among treatments. The control treatment showed higher CMCase activity when compared with the treatments containing supplemental sources of nitrogen. However, urea supplementation provided higher CMCase activity compared to albumin.

A comparative study of β-1, 4-endoglucanase (possessing β-1, 4-exoglucanase activity) from Bacillus subtilis LH expressed in Pichia pastoris GS115 and Escherichia coli Rosetta (DE3)

β-1, 4-Endoglucanase (EG) from Bacillus subtilis LH was expressed in Escherichia coli Rosetta (DE3) and Pichia pastoris GS115, respectively. The CMCase activity of EG (EGE) from the cell lysate of DE3 reached 20,010U/ml, and that of EG (EGP) from the supernatant of GS115 was only 2008U/ml. EGE and EGP were bifunctional cellulases excluding β-1, 4-glucosidase (BGL). The CMCases of them, optimally active at 65°C and pH 6.8, exhibited more than 80% residual activity at pH 5–10 and 60% activity at 40–70°C and pH 5–9. EGE (EGP) mixed with BGL had more than 1.5-fold higher CMCase and filter paperase activities compared to EGE (EGP). N-glycosylation protected EGP from immobilized-papain attack and accounted for 30kDa and a higher thermostability, whereas EGE was decomposed into a 33kDa active truncated EG (EGT) and two 18kDa fragments. EGE and EGP performed much better than EGT in denim biostoning.

Improvement of carboxymethyl cellulase production from Chaetomium cellulolyticum NRRL 18756 by mutation and optimization of solid state fermentation

The purpose of the present investigation was to enhance the production of the industrially important enzyme carboxymethyl cellulase (CMCase) by subjecting the wild cellulase producing fungal strain Chaetomium cellulolyticum NRRL 18756 to various doses of gamma irradiation (0.25, 0.5, 0.75, 1.0, 1.25, 1.5, 1.75 and 2.0 KGy). Among all the mutants tested, M-7 obtained at 0.5 KGy of C. cellulolyticum strain showed highest extracellular CMCase production in a yield 1.6-fold exceeding that of the wild type. Optimal conditions for the production of CMCase by the mutant fungal strain using solid-state fermentation were examined. The optimized medium consisted of sugarcane bagasse supplemented with 1% (w/w) peptone, 2.5 mM MgSO4, and 0.05% (v/w) Tween 80. Optimal moisture content and initial pH was 40% (v/w) and 5.0 to 6.5, respectively. The medium was fermented at 40°C for 4 days. The resulting CMCase yield was 4.0-fold more than that of the wild type strain grown on the basal wheat bran medium. Some characteristics of partially purified CMCase from the mutant and wild type of C. cellulolyticum were investigated. The partially purified mutant CMCase was more stable than the wild type CMCase. Thus, the higher thermostability of mutant CMCase makes it a potential candidate for commercial and industrial process. Key word: Chaetomium cellulolyticum, carboxymethyl cellulase, mutation, optimization, solid state fermentation, characterization.

토양으로부터 분리한 Bacillus licheniformis KJ 9의 특성

고품질의 발효퇴비를 생산할 목적으로 자연계로부터 효소활성이 높고 식물병원성 균주에 대한 항균활성이 우수한 미생물을 순수분리 하여 배양학적 특성을 검토하였다. 부엽토 등으로부터 CMCase, protease 및 chitinase 활성이 높고 식물병원성 균주에 대하여 항균활성이 우수한 KJ-9균주를 분리한 다음 형태학적 및 생화학적 특성을 검토하고 Bergey's Mannual of Systematic Bacteriology의 방법과 ATB (Automated Identification) computer system (Bio Merieux, France)을 이용한 API 50 CHL Carbohydrate Test Kit(Bio Merieux, France)를 통하여 동정한 결과 Bacillus licheniformis로 밝혀졌다. B. licheniformis KJ-9의 최적배지 성분을 검토한 결과 탄소원은 1.5% soluble starch, 질소원은 0.5% yeast extract, 무기염은 0.05% <TEX>$MgSO_4{\cdot}7H_2O$</TEX> 이었으며, 배지의 초기 pH는 7.0, 배양온도는 <TEX>$50^{\circ}C$</TEX> 그리고 진탕속도는 180 rpm으로 밝혀졌다. 최적배양 조건으로 B. licheniformis KJ-9를 배양하였을 때 36~60 hr째의 배양액은 잿빛 곰팡이병을 유발하는 Botrytis cinerea, 잎마름병의 원인균인 Corynespora cassicola, 시들음병을 유발하는 Fusarium oxysporum, 잘록병의 원인균인 Rhizocfonia solani의 식물병원성 미생물 균사생장을 효과적으로 억제하였다. In order to produce high-quality fermenting composts, a microorganism was isolated from the natural world. The bacterium has not only in high enzyme activities but also had good antimicrobial activities against phytopathogenic microorganisms. Its cultivating characteristics were then investigated. Bacterium KJ-9, which contains high CMCase, protease and chitinase activities and excellent antimicrobial activities against phytopathogenic microorganisms, was separated from leaf mold and identified as Bacillus licheniformis by two methods: Bergey's Manual of Systematic Bacteriology and API 50 CHL Carbohydrate Test Kit (Bio Merieux, France) using an ATB (Automated Identification) computer system (Bio Merieux, France). Optimal medium for cultivation of B. licheniformis was 2% soluble starch as a carbon source, 0.5% yeast extract as a nitrogen source and 0.05% <TEX>$MgSO_4{\cdot}7H_2O$</TEX>. Optimal growth conditions of pH, temperature and shake speed were pH 7.0, <TEX>$50^{\circ}C$</TEX> and 180 rpm, respectively. Culture broth of B. licheniformis KJ-9 cultured for 36~60 hr was effective in fungicidal activities against plant pathogens including Botrytis cinerea, Corynespora cassicola, Fusarium oxysporum, and Rhizoctonia solani.

Yield performances and changes in enzyme activities of Pleurotus spp. ( P. ostreatus and P. sajor-caju) cultivated on different agricultural wastes

Pleurotus ostreatus and Pleurotus sajor-caju were studied for their ability to produce laccase and carboxymethylcellulase (CMCase) enzymes on different agricultural wastes under solid state fermentation. The spawns of P. ostreatus and P. sajor-caju were inoculated on different agricultural wastes including viticulture wastes, wheat straw, paddy straw, sesame straw, sawdust as well as the mixtures of these wastes with wheat bran. The carbon and nitrogen contents of substrates containing bran were the highest. The laccase activities of P. ostreatus and P. sajor-caju reached the highest values on the day 10 of mycelial growth. This enzyme activity was higher on the substrates containing bran which had high nitrogen and low C/N ratio than the other tested substrates with no bran. The CMCase activities of P. sajor-caju and P. ostreatus had two peaks on the 5th day of mycelial growth and after first flash. P. ostreatus and P. sajor-caju grown on substrates containing wheat bran had higher biological efficiencies and total yields as well as higher CMCase and laccase activities.

Carboxymethylcellulase production by the anaerobic rumen fungusNeocallimastix sp. GMLF7

Anaerobic fungi have highly active fibrolytic enzymes and these enzymes are attractive for scientific research. We isolated a ruminal fungus of the genusNeocallimastix sp., named GMLF7, which could survive on a variety of cellulosic material such as carboxymethylcellulose (CMC), fibrous cellulose, avicel and wheat straw. Carboxymethylcellulase production was investigated with the above carbon sources and high CMCase activity was obtained with CMC (73.75 U/ml), fibrous cellulose (72.68 U/ml) and avicel (70.03 U/ml). While growth temperature of the microorganism was 39°C, for CMCase activity optimal temperature was 50°C and optimal pH 6.5. Enzyme reached maximum activity in 60–180 min at 50°C. Substrate concentration also affected the enzyme activity which was increased more than 2 fold with 40 mg/ml CMC. Activity was measured in presence of various divalent ions and reagents, and Co2+ and DTT has a positive effect on the enzyme activity.

Intra-strain protoplast fusion enhances carboxymethyl cellulase activity in Trichoderma reesei

Protoplasts were isolated from Trichoderma reesei strain PTr2 using Lysing enzymes (Sigma Chemicals Co., USA) with 0.6 M KCl as osmotic stabilizer. Intra-strain protoplast fusion has been carried out using 40% polyethylene glycol with STC (sorbitol, Tris–HCl, CaCl 2) buffer. The fused protoplasts of T. reesei have been regenerated on carboxymethyl cellulose agar (CMCA) selective medium and 15 fusants were selected for further studies. Most of the fusants exhibited fast mycelial growth and abundant sporulation on PDA as compared to non-fusant and parent strains. Prominent clearing zones that appeared around the mycelial growth of most of the fusant progenies when grown on CMCA indicated the secretion of high level extracellular carboxymethyl cellulase (CMCase) than the non-fusants and parent. High CMCase activity was estimated with 80% of the fusants and more than two-fold increase in enzyme activity was recorded with two fusants, SFTr2 and SFTr3 as compared to the parent strain PTr2. Results of the present study demonstrated the scope and significance of the protoplast fusion technique, which can be used to develop superior hybrid strains of filamentous fungi that lack inherent sexual reproduction.

Isolation and characterization of mesophilic cellulose-degrading bacteria from flower stalks-vegetable waste co-composting system

Fifteen mesophilic bacteria with high C(x) cellulase activities were isolated and purified from a mixed-culture enriched from a flower stalks-vegetable waste co-composting system. A CMCase test showed that the enzyme activity of these isolates ranged from 7.9 to 28.0 U ml(-1). Although filter paper degrading capability was low in single culture, significant synergetic cellulose degradation were detected in four groups of mixed cultures, their degradation rates were 23.5%, 26.3%, 19.4% and 24.5%, respectively. Study of morphological and physiological characters of five predominant isolates which possess high CMCase and had positive effect on synergetic cellulose degradation in mixed culture system showed that two of them were closely related to Bacillus pasteurii and Bacillus cereus, whereas the rest belong to the genus Halobacillus, Aeromicrobium and Brevibacterium, respectively.

OPTIMIZATION OF CELLULASE PRODUCTION WITH PENICILLIUM NALGIOVENSE Sll GROWN ON PRETREATED WHEAT POLLARD

The cellulase production with Penicillium nalgiovense S11 on wheat pollard was enhanced using substrate pretreatments, i.e.: (i) mechanic process by Wiley milling, (ii) reducing sugars removal by water soaking, and (iii) chemical pretrcatment by 0.5% NaOH soaking at 100°C. The enzyme production stated as enzyme activities of all prctrcated substrates were higher than the untreated substrate. Although soaking with water showed significant increase in enzyme activities, the highest CMCase (EC 3.2.1.4), FPase (filter papcrase) and p- glucosidase (EC 3.2.1.21) were observed on NaOH pretreated pollard. The NaOH prctreatment also enhanced the enzyme production by increasing substrate concentration from 2 to 4%. The optimal incubation time in the cellulase production on 4% NaOH-pretrcated pollard was observed on the fifth day. Addition of 250 ppm glucose also increased the enzyme activities. The optimal treatments increased the specific activities of CMCase, FPase, and |3-glucosidase into 60, 4, and 198 times, respectively, as compared to the specific activities on 2% unpretreated pollard. Keywords : Cellulase production/ pollard pretreatments/ water soaking/ NaOH soaking

SYNERGISTIC ACTIVITY OF ENZYMES PRODUCED BY EUPENICILLIUM JAVANICUM AND ASPERGILLUS NIGER NRRL 337 ON PALM OIL FACTORY WASTES

The use of palm kernel cake (PKC) and palm oil mill effluent (POME), substances from palm oil factory wastes, for monogastric is limited by their high cellulose and mannan contents. Hydrolytic enzymes have been supplemented to increase the nutrient digestibility. The maximal digestibility was obtained in the synergistic action of all enzyme components including B-D-endoglucanase (CMCase), B-D-glucosidase, B-D-mannanase, p-D-mannosidase, and oc-D- galactosidase. Two kinds of enzymes produced by Eupenicillium javanicum and Aspergillus niger NRRL 337 on the submerged culture containing 3% coconut meal were selected to hydrolyze PKC or dry POME. Enzyme from E. javanicum contained higher CMCase, B-D-mannanase, and a-D-galactosidase activities, while that from A. niger NRRL 337 contained more p-D-glucosidase and p-D-mannosidase activities. Saccharification (hydrolytic) activities of enzyme mixtures on PKC and POME were determined at pH 5.0, the optimal pH for p-D-mannanase from E. javanicum, and at 5.4 the optimal pH for a-D-galactosidase from E. javanicum and P-D-glucosidase from A. niger NRRL 337. The enzyme proportions of E. javanicum and A. niger NRRL 337 were 100 : 0, 80 : 20, 60 : 40, 40 : 60, and 0 : 100%. The highest Saccharification activity on both substrates was observed on the mixture of 80% A. niger NRRL 337. The pH levels did not significantly affect Saccharification activity. Fiber components in PKC were more digestable than in POME. Further analysis on the reducing sugar components using thin layer chromatography showed that more monomers were produced in the 60 or 80% of A. niger NRRL 337. The glycosidases of A. niger NRRL 337 played more important role in the Saccharification activity. Keywords: Synergistic activity/ palm kernel cake/palm oil mill effluent/ Eupenicillium javanicum/ Aspergillus niger NRRL 337

A cell surface display system using novel GPI-anchored proteins in Hansenula polymorpha.

A cell surface display system was developed in yeast Hansenula polymorpha. The four genes HpSED1, HpGAS1, HpTIP1and HpCWP1, encoding glycosylphosphatidyl-inositol (GPI)-anchored cell surface proteins from H. polymorpha, were cloned, characterized and evaluated for their efficacies as cell surface display motifs of reporter proteins. Sequence analysis of these genes revealed that each encodes a typical GPI-anchored protein that is structurally similar to a counterpart gene in S. cerevisiae. The genes showed a high content of serine-threonine (alanine) and harboured a putative secretion signal in the N-terminus and the GPI-attachment signal in the C-terminus. The surface anchoring efficiency of these putative cell surface proteins was tested by fusion to the C-terminal of carboxymethylcellulase (CMCase) from Bacillus subtilis. In all cases, high CMCase activities were detected in intact cell fraction, indicating anchoring of CMCase to the cell surface. HpCwp1p, HpGas1p and the 40 C-terminal amino acids of HpTip1p from H. polymorpha exhibited a comparatively high CMCase surface anchoring efficiency. When these proteins were used as anchoring motifs for surface display of the glucose oxidase (GOD) from Aspergillus niger, most enzyme activity was detected at the cell surface. Fluorescence activated cell sorter (FACS) analysis of cells displaying GOD on the cell surface demonstrated that GOD was well exposed on the cell surface. HpCwp1p showed the highest anchoring efficiency among others.

Isolation and characterization of a lichenan-degrading hydrophobic endoglucanase of Clostridium thermocellum

Endoglucanase 7 (EG7) of Clostridium thermocellum was isolated from a recombinant strain of Escherichia coli TG1 cells harbouring recombinant plasmid pCU110 containing the cel7 gene of C. thermocellum. The enzyme was purified to electrophoretic homogeneity and was presented as two components with a molecular mass of 49 and 47 kDa and a pI of 4.35 and 4.30, respectively. The enzyme was shown to have optimum pH of 5.5 and optimum temperature of 55–60° C with carboxymethylcellulose (CMC) as a substrate. EG7 displayed hyper-lichenase, high CMCase, low cellobiosidase and negligibly small activities towards Avicel, amorphous cellulose, laminarin and xylan. The enzyme was shown to be stable at 55° C and within a broad range of pH from 4.5 to 11.0. It is insensitive towards ethanol (up to 5%) and end-product (cellobiose or glucose) inhibition. The hydrophobic nature of the protein, as revealed by retarded elution on gel filtration columns, resulted in an unprecedentedly high yield (about 80%) of purified enzyme. Due to the above-mentioned characteristics, the enzyme should to be quite suitable for use in the mashing process of beer brewing.

The effect of sucrose supplements on particle-associated carboxymethylcellulase (EC 3.2.1.4) and xylanase (EC 3.2.1.8) activities in cattle given grass-silage-based diet.

Carboxymethylcellulase (EC 3.2.1.4; CMCase) and xylanase (EC 3.2.1.8) activities were assayed in rumen fluid and from microbes closely associated either with rumen particulate material or with feed particles incubated in nylon bags in the rumen of cattle. The cattle were fitted with a permanent rumen cannula and a simple 'T'-piece duodenal cannula and were given four diets in a 4 x 4 Latin Square experiment. The basal diet (diet C) consisted of grass silage, barley and rapeseed meal (700, 240 and 60 g/kg total dry matter (DM)) given at the rate of 5.3 kg/d or supplemented with 1.0 kg sucrose/d given twice daily (diet S), twice daily with 0.25 kg sodium bicarbonate/d (diet B) or as a continuous intrarumen infusion (diet I). Giving sucrose supplements decreased CMCase and xylanase activities extracted from microbes associated with rumen particulate material or feed particles incubated in nylon bags as compared with diet C. Supplementation of the sucrose diet with sodium bicarbonate resulted in higher CMCase and xylanase activities than other sucrose diets (S and I). Particle-associated CMCase and xylanase activities were found to be very sensitive in detecting differences in the rumen environment and were related to changes in cell wall digestion. The activities were highly correlated with disappearance of DM and neutral-detergent fibre from nylon bags incubated in the rumen, rumen and total digestion of cell-wall carbohydrates and rumen pool size of cell-wall carbohydrates. It was concluded that the attachment of fibrinolytic enzymes is involved in the depression of fibre digestion. Particle-associated CMCase and xylanase activities were much higher when measured from rumen particulate material than from feed particles incubated in nylon bags.

Cellulase production by halophytic fungi: II. The role of some cultural conditions on cellulase production by Ulocladium chartarum

Summary Cellulase activities of Ulocladium chartarum were found to be influenced by the mode of incubation as well as by the age of the fungal culture. Maximal cellulase activities were, however, maintained after 3 weeks using the surface culture mode of incubation. U. chartarum was grown at different temperatures, where maximal CMCase and FPase activities were achieved at 30°C. The cellulase activities were also tested using initially adjusted p H as well as buffered media. The highest CMCase and FPase activities (0.289 and 0.043 unit/ml respectively) were obtained with p U 6.5 and the phosphate buffer gave better results as compared with the acetate buffer.

Transfer of DNA coding for cellulases from Cellulomonas species to Bacillus subtilis by protoplast fusion

Cellulose utilising hybrids between Cellulomonas spp. and Bacillus subtilis were isolated after PEG mediated protoplast fusion. These stable hybrids selected using genomic markers harboured DNA from both the parents. Higher CMCase and β-glucosidase activities were detected extracellularly in case of one of the hybrids unlike only CMCase in case of Cellulomonas.