Parametric Optimization of Cultural Conditions for Carboxymethyl Cellulase Production Using Pretreated Rice Straw by Bacillus sp. 313SI under Stationary and Shaking Conditions

Carboxymethyl cellulase (CMCase) provides a key opportunity for achieving tremendous benefits of utilizing rice straw as cellulosic biomass. Out of total 80 microbial isolates from different ecological niches one bacterial strain, identified as Bacillus sp. 313SI, was selected for CMCase production under stationary as well as shaking conditions of growth. During two-stage pretreatment, rice straw was first treated with 0.5 M KOH to remove lignin followed by treatment with 0.1 N H2SO4 for removal of hemicellulose. The maximum carboxymethyl cellulase activity of 3.08 U/mL was obtained using 1% (w/v) pretreated rice straw with 1% (v/v) inoculum, pH 8.0 at 35°C after 60 h of growth under stationary conditions, while the same was obtained as 4.15 U/mL using 0.75% (w/v) pretreated substrate with 0.4% (v/v) inoculum, pH 8.0 at 30°C, under shaking conditions of growth for 48 h. For maximum titre of CMCase carboxymethyl cellulose was optimized as the best carbon source under both cultural conditions while ammonium sulphate and ammonium nitrate were optimized as the best nitrogen sources under stationary and shaking conditions, respectively. The present study provides the useful data about the optimized conditions for CMCase production by Bacillus sp. 313SI from pretreated rice straw.

Download Full-text

Production and Optimization of Cellulase Activity of Thermomonospora Viridis Isolated From Rice Straw

Bangladesh Journal of Botany ◽

10.3329/bjb.v50i2.54097 ◽

2021 ◽

Vol 50 (2) ◽

pp. 395-404

Author(s):

Faozia Faleha Sadida ◽

Ma Manchur

Keyword(s):

Enzyme Activity ◽

Metal Ions ◽

Rice Straw ◽

Cellulase Activity ◽

Nitrogen Sources ◽

Cellulase Production ◽

Tetraacetic Acid ◽

Carbon And Nitrogen Sources ◽

Carbon And Nitrogen ◽

Optimum Ph

A highly cellulolytic actinomycete SR1 was locally isolated from rice straw and provisionally identified as Thermomonospora viridis. Optimum pH, temperature, carbon and nitrogen sources for its cellulase production were 6.5, 35°C, Carboxymethyl cellulase (CMC) and yeast extract, respectively whereas those of cellulase activity were 7.5, 40°C, CMC and peptone respectively. The effects of various metal ions and different reductant and inhibitors on its cellulase activity were investigated. Univalent Ag+ was found to decrease the enzyme activity whereas increased by bivalent Mg2+. Ethylene diamine tetraacetic acid (EDTA) caused remarkable decrease of cellulase activity but β-Mercaptoethanol stimulated its cellulase activity. Bangladesh J. Bot. 50(2): 395-404, 2021 (June)

Download Full-text

Optimization of cellulase production by bacillus sp. BPPT CC RK2 with pH and temperature variation using response surface methodology

E3S Web of Conferences ◽

10.1051/e3sconf/20186702051 ◽

2018 ◽

Vol 67 ◽

pp. 02051

Author(s):

Misri Gozan ◽

Andre Fahriz Harahap ◽

Chandra Paska Bakti ◽

Siswa Setyahadi

Keyword(s):

Response Surface Methodology ◽

Response Surface ◽

Agricultural Waste ◽

Cellulase Activity ◽

Preliminary Test ◽

Cellulase Production ◽

Bacillus Sp ◽

Positive Effects ◽

Rsm Optimization ◽

Quadratic Effects

Indonesia has abundant ethanol biomass feedstocks. However the second-generation ethanol production process is still hampered by the unavailability of cellulase enzyme in the process of decomposition of lignocellulose into saccharides that can be processed into ethanol through fermentation. Cellulase is known as exozyme produced by Bacillus sp. in submerged fermentation. In this study, cellulase production by Bacillus sp. CC BPPT RK2 on natural and abundant agricultural waste substrates (rice bran and coconut water) was evaluated by investigating the optimum conditions for cellulase production in a 50 ml laboratory scale. Preliminary test using Luria Bentani (LB) medium with additional CMC (1%) were done to select optimum range of pH and Temperature. The preliminary tests results were then followed by optimization of pH and temperature, which were carried out using response surface methodology (RSM). RSM optimization model showed optimum values 6.23 for pH and 40.04 °C, with 14 terms (each with 1 degree of freedom), 4 linear effects, 6 interaction effects and 4 quadratic effects. These optimization by RSM results were slightly different compared to preliminary test, showing the effect of interactions between parameters. The characteristics of interaction among variables tested against the cellulase activity are reported in this study including: positive effects on cellulase activity of the resulting responses; negative interactions affecting the response of cellulase activity; synergistic interaction; and antagonistic interactions between each other.

Download Full-text

OPTIMIZATION OF CELLULASE ENZYME PRODUCTION FROM Aspergillus oryzae FOR INDUSTRIAL APPLICATIONS

World Journal of Biology and Biotechnology ◽

10.33865/wjb.002.02.0088 ◽

2017 ◽

Vol 2 (2) ◽

pp. 155 ◽

Cited By ~ 1

Author(s):

Hassan Sher ◽

Muhammad Faheem ◽

Abdul Ghani ◽

Rashid Mehmood ◽

Hamza Rehman ◽

...

Keyword(s):

Enzyme Production ◽

Time Course ◽

Value Added ◽

Cellulase Production ◽

Industrial Applications ◽

Inoculum Size ◽

Cellulosic Biomass ◽

High Yield ◽

Carboxymethyl Cellulase ◽

Cellulase Enzyme

Cellulases are the hydrolytic group of enzymes, responsible for release of sugars in the bioconversion of the cellulosic biomass into a variety of value added industrial products. Fungal isolated cellulases are well studied and playing a significant role in various industrial processes. Enzymatic depolymerisation of cellulosic material has been done by the various fungal isolated enzymes. In the present study, the cultivation conditions for cellulase production from Aspergillus species were optimized. Optimization of scarification conditions such as time course, inoculum size, carbon source and concentration, nitrogen source, various pH levels were performed for the production of extracellular carboxymethyl cellulase and endoglucanase enzyme. The result exhibited, 15 % inoculums size, corncobs 2 % concentration, Urea and medium pH 7 at 30oC supported high yield of carboxymethyl cellulase (38.80 U/ml/min) and exoglucanase enzyme (10.94 U/ml/min) through a submerged fermentation (SmF). In future biotechnological applications in cellulase enzyme production attain a vital role to obtain high degradable yield.

Download Full-text

Extracellular cellulase production by tropical isolates of Aureobasidium pullulans

Canadian Journal of Microbiology ◽

10.1139/w05-053 ◽

2005 ◽

Vol 51 (9) ◽

pp. 773-776 ◽

Cited By ~ 23

Author(s):

T Kudanga ◽

E Mwenje

Keyword(s):

Carboxymethyl Cellulose ◽

Cell Walls ◽

Aureobasidium Pullulans ◽

Plant Cell Wall ◽

Synthetic Medium ◽

Cellulase Activity ◽

Nitrogen Sources ◽

Cellulase Production ◽

Cellulolytic Activity ◽

Specific Activities

Cellulase production by Aureobasidium pullulans from the temperate regions has remained speculative, with most studies reporting no activity at all. In the current study, tropical isolates from diverse sources were screened for cellulase production. Isolates were grown on a synthetic medium containing cell walls of Msasa tree (Brachystegia sp.) as the sole carbon source, and their cellulolytic activities were measured using carboxymethyl cellulose and α-cellulose as substrates. All isolates studied produced carboxymethyl cellulase (endoglucanase) and alpha-cellulase (exoglucanase) activity. Endoglucanase-specific activities of ten selected isolates ranged from 2.375 to 12.884 µmol glucose·(mg protein)–1·h–1, while activities on α-cellulose (exoglucanase activity) ranged from 0.293 to 22.442 µmol glucose·(mg protein)–1·day–1. Carboxymethyl cellulose induced the highest cellulase activity in the selected isolates, while the isolates showed variable responses to nitrogen sources. The current study indicates that some isolates of A. pullulans of tropical origin produce significant extracellular cellulolytic activity and that crude cell walls may be good inducers of cellulolytic activity in A. pullulans.Key words: Aureobasidium pullulans, plant cell wall, cellulases, endoglucanase, exoglucanase.

Download Full-text

OPTIMASI NUTRISI MEDIA PERTUMBUHAN BAKTERI TERMOFIL PENGHASIL SELULASE DARI SUMBER AIR PANAS RIMBO PANTI

Jurnal Riset Kimia ◽

10.25077/jrk.v5i1.165 ◽

2015 ◽

Vol 5 (1) ◽

pp. 1

Author(s):

Armaini ◽

Abdi Dharma ◽

Sumaryati Syukur ◽

Jamsari

Keyword(s):

Nitrogen Source ◽

Hot Springs ◽

Inorganic Nitrogen ◽

Carbon Sources ◽

Cellulase Activity ◽

Nitrogen Sources ◽

Cellulase Production ◽

Optimum Concentration ◽

Crystalline Cellulose ◽

Inorganic Nitrogen Source

Optimization have been done on the media for the growth of the isolated thermophiles bacteria from hot springs Rimbo Panti, the nutrients comprising variety of carbon sources such as CMC (carboxymethyl cellulose), avicell (micro crystalline cellulose), and cellobiose, with a variety of sources organic nitrogen, peptone, extracts yeast, tryptone, and urea, as well as variations consist of inorganic nitrogen sources, KNO3, NaNO3, (NH4)2SO4, and (NH4)NO3. Determination of cellulase activity performed using DNS reagent (3,5-dinitro salicylic acid). Maximum cellulase production with high activity based on the results of this research, the best of carbon source is CMC with optimum concentration 0.125%, inorganic nitrogen source is peptone with the optimum concentration of 0.3 to 0.4% and the inorganic nitrogen source is (NH4)2SO4 with optimum concentration of 0.2 - 0.25%. Optimization of size of inoculums obtained the optimum amount of inoculums 2%. Keywords: Optimization, thermophiles bacteria, cellulose, carbon sources, nitrogen sources

Download Full-text

Carboxymethyl cellulase production from a Paenibacillus sp.

Hemijska industrija ◽

10.2298/hemind150222038m ◽

2016 ◽

Vol 70 (3) ◽

pp. 329-338 ◽

Cited By ~ 2

Author(s):

Katarina Mihajlovski ◽

Sladjana Davidovic ◽

Milica Carevic ◽

Neda Radovanovic ◽

Slavica Siler-Marinkovic ◽

...

Keyword(s):

Culture Supernatant ◽

Soluble Sugars ◽

Cellulase Activity ◽

Cellulase Production ◽

Carboxymethyl Cellulase ◽

Carbon And Nitrogen ◽

Cmcase Activity ◽

Working Volume ◽

Paenibacillus Sp ◽

Initial Ph

Cellulases are industrially important enzymes with a potential to convert cellulose into fermentable sugars. Novel bacterial isolate Paenibacillus sp. CKS1 was tested for cellulase activity and the optimal conditions for carboxymethyl cellulase (CMCase) production were determined. Maximum CMCase activity was obtained in the third passage of the bacterial culture after 3 days of incubation at 30?C. Cellobiose and yeast extract was the optimal source of carbon and nitrogen for induction of CMCase activity. In addition, with initial pH 7 of the medium and 40 ml of working volume in 500 ml culture flasks with shaking at 150 rpm, the maximum CMCase activity in a crude culture supernatant reached value of 0.532?0.006 U/ml. For crude CMCase, optimal temperature was 50?C and optimal pH 4.8, respectively. HPLC analysis confirmed the bacterium is capable to hydrolise CMC to glucose and other soluble sugars.

Download Full-text

Isolation, identification and in silico study of native cellulase producing bacteria

Current Proteomics ◽

10.2174/1570164617666191127142035 ◽

2019 ◽

Vol 17 ◽

Author(s):

Farzane Kargar ◽

Mojtaba Mortazavi ◽

Mahmood Maleki ◽

Masoud Torkzadeh Mahani ◽

Younes Ghasemi ◽

...

Keyword(s):

Growth Rate ◽

Bacillus Cereus ◽

16S Rdna ◽

Enzyme Production ◽

Rational Design ◽

Cellulase Production ◽

Bacillus Species ◽

Bacillus Sp ◽

Chain Polymer ◽

Bacillus Toyonensis

Aims: The purpose of this study was to screen the bacteria producing cellulase enzymes and their bioinformatics studies. Background: Cellulose is a long-chain polymer of glucose that hydrolyzes by cellulases to glucose molecules. In order to design the new biotechnological applications, some strategies have been used as increasing the efficiency of enzyme production, generating cost-effective enzymes, producing stable enzymes and identification of new strains. Objective: On the other hand, some bacteria special features have made them suitable candidates for the identification of the new source of enzymes. In this regard, some native strains of bacteria were screened. Method: These bacteria were grown on a culture containing the liquid M9 media containing CMC to ensure the synthesis of cellulase. The formation of a clear area in the culture medium indicated decomposition of cellulose. In the following, the DNA of these bacteria were extracted and their 16S rDNA genes were amplified. Result: The results show that nine samples were able to synthesize cellulase. In following, these strains were identified using 16S rDNA. The results show that these screened bacteria belonged to the Bacillus sp., Alcaligenes sp., Alcaligenes sp., and Enterobacter sp.conclusionThe enzyme activity analysis shows that the Bacillus toyonensis, Bacillus sp. strain XA15-411 Bacillus cereus have produced the maximum yield of cellulases. However, these amounts of enzyme production in these samples are not proportional to their growth rate. As the bacterial growth chart within 4 consecutive days shows that the Alcaligenes sp. Bacillus cereus, Bacillus toyonensis, Bacillus sp. strain XA15-411 have a maximum growth rate. The study of the phylogenetic tree also shows that Bacillus species are more abundant in the production of cellulase enzyme. These bioinformatics analyses show that the Bacillus species have different evolutionary relationships and evolved in different evolutionary time. Other: However, for maximum cellulase production by this bacteria, some information as optimum temperature, optimum pH, carbon and nitrogen sources are needed for the ideal formulation of media composition. The cellulase production is closely controlled in microorganisms and the cellulase yields appear to depend on a variety of factors. However, the further studies are needed for cloning, purification and application of these new microbial cellulases in the different commercial fields as in food, detergent, and pharmaceutical, paper, textile industries and also various chemical industries. However, these novel enzymes can be further engineered through rational design or using random mutagenesis techniques.

Download Full-text

Production and optimization of cellulase from agricultural waste and its application in bioethanol production by simultaneous saccharification and fermentation

Management of Environmental Quality An International Journal ◽

10.1108/meq-07-2015-0128 ◽

2016 ◽

Vol 27 (1) ◽

pp. 22-35 ◽

Cited By ~ 2

Author(s):

Elsa Cherian ◽

M. Dharmendira Kumar ◽

G. Baskar

Keyword(s):

Design Methodology ◽

Simultaneous Saccharification And Fermentation ◽

Agricultural Waste ◽

Cellulase Production ◽

Reducing Sugar ◽

Bioethanol Production ◽

Corn Cob ◽

Content Type ◽

Optimized Conditions ◽

Simultaneous Saccharification

Purpose – The purpose of this paper is to optimize production of cellulase enzyme from agricultural waste by using Aspergillus fumigatus JCF. The study also aims at the production of bioethanol using cellulase and yeast. Design/methodology/approach – Cellulase production was carried out using modified Mandel’s medium. The optimization of the cellulase production was carried out using Plackett-Burman and Response surface methodology. Bioethanol production was carried out using simultaneous saccharification and fermentation. Findings – Maximum cellulase production at optimized conditions was found to be 2.08 IU/ml. Cellulase was used for the saccharification of three different feed stocks, i.e. sugar cane leaves, corn cob and water hyacinth. Highest amount of reducing sugar was released was 29.1 gm/l from sugarcane leaves. Sugarcane leaves produced maximum bioethanol concentration of 9.43 g/l out of the three substrates studied for bioethanol production. Originality/value – The present study reveals that by using the agricultural wastes, cellulase production can be economically increased thereby bioethanol production.

Download Full-text

Optimization of cultural conditions affecting improved bioactive metabolite production by endophytic fungus Trichoderma harzianum

Bionatura ◽

10.21931/rb/2021.06.04.8 ◽

2021 ◽

Vol 6 (4) ◽

pp. 2187-2192

Author(s):

Rashid Rahim Hateet ◽

Zainab Alag Hassan ◽

Abdulameer Abdullah Al-Mussawi ◽

Shaima Rabeea Banoon

Keyword(s):

Antibacterial Activity ◽

Trichoderma Harzianum ◽

Endophytic Fungus ◽

Nitrogen Sources ◽

Bioactive Metabolites ◽

Bioactive Metabolite ◽

Metabolite Production ◽

Surface Sterilization ◽

Cultural Conditions

The present study aimed to optimize cultural conditions for optimum bioactive metabolite production by endophytic fungus Trichoderma harzianum, isolated by surface sterilization method from the leaf of the eucalyptus plant. The fungus was identified based on morphological characterization. Fungal metabolites were carried out by ethyl acetate solvent. The antibacterial activity was tested against Escherichia coli (ATCC 25922) and Staphylococcus aureus (NCTC 6571). Various carbon, nitrogen sources, pH, temperature, incubation period, and NaCl on the antibacterial metabolite production were studied. Bioactive metabolite production of T. harzianum exhibits a broad spectrum of in vitro antibacterial activity against two strains of bacteria. For the optimum production of bioactive metabolites, Dextrose and Glucose were found to be the best sources of carbon and the best sources of Nitrogen Yeast extract (YE) and (NH4)2SO. The maximum production of bioactive metabolites occurs at pH 7 and 25°C.; the NaCl showed a positive influence on bioactive metabolites.

Download Full-text

Wild type Bacillus subtilis treated with ethyl methyl sulphonate produced catabolite insensitive mutants with improved cellulase production

10.21203/rs.3.rs-278081/v1 ◽

2021 ◽

Author(s):

Oladipo Olaniyi

Keyword(s):

Bacillus Subtilis ◽

Carbon Source ◽

Enzyme Production ◽

Cellulase Activity ◽

Cellulase Production ◽

Minimal Salt Medium ◽

Salt Medium ◽

Production Medium ◽

Wild Type ◽

Ethyl Methyl

Abstract The goal of this present investigation was to mutagenize Bacillus subtilis with Ethyl Methyl Sulphonate (EMS), screen the mutants for cellulase production and evaluate the influence of different glucose concentrations on their cellulase production potentials. The wild type B. subtilis was treated with 20, 40, 60 and 80 µl of EMS and the mutants generated were screened for cellulase production in minimal salt medium containing carboxylmethylcellulose (CMC) as the carbon source. Quantitatively, cellulase activity and protein contents were determined by dinitrosalicylic acid and Lowry methods respectively. Seven mutants were developed from each of the EMS concentration bringing the total to twenty-eight from all the concentrations. Approximately 14 and 57% of the mutants developed from 40 and 60µl of EMS had higher cellulase activities than the wild type, while none of the mutants developed from 20 and 80 µl of EMS had better activities than the wild type. The supplementation of 0.2, 0.5, 1.0 and 1.5% glucose in enzyme production medium caused approximately 100, 14, 29 and 14% cellulase repression respectively in the mutants developed from 60µl EMS. Mutants MSSS02 and MSSS05 were considered as catabolite insensitive mutants because their cellulase production were enhanced in comparison to wild type.

Download Full-text