Cellulase Enzyme Production Research Articles

Bioethanol production from groundnut shell using microorganisms derived from sugarcane root soil

Bioethanol is a form of renewable energy that can be produced from fuel or energy crops. Ethanol is produced by the fermentation of sugars present in agricultural feed stocks and crop residues. This study investigates the use of agro wastes like groundnut shells for ethanol production. Initially, the groundnut shells were washed, dried and grinded into powder. Then it was subjected to ethanol production using yeast. After 20 days of incubation the ethanol was estimated using potassium dichromate method. The maximum ethanol yield (1.55%) was obtained when 1% yeast was used. To increase the efficiency of ethanol production, the cellulolytic bacteria was isolated from the cow dung dumped soil site. The 10 bacterial isolates were screened for cellulase enzyme production. Among them one bacterium showed maximum decolourization of congored which was subjected for enzyme production in nutrient broth. The organism showed maximum enzyme activity of 558.12 U/ml. The isolated cellulolytic bacteria were identified as Bacillus anthracis using 16s rDNA sequencing. Ethanol production from the groundnut shells was again carried out with various concentration of crude cellulase enzyme isolated from the bacteria. The estimation result showed 3.8% of ethanol as maximum. Then the ethanol was condensed using a rotary evaporator and when again estimated, showed 7.3% of ethanol. Finally, the presence of ethanol was confirmed by iodoform test. Thus, groundnut shells can efficiently be used for the production of ethanol, which can be used as a high potential fuel source for transportation in the future.

Cellulase Enzyme Production Using Actinobacillus sp. on Several Alternative Growth Media

Cellulase enzymes are widely used in the food, beverage, animal feed, textile, and paper industries. The high cost of producing enzymes and low enzyme activity provides opportunities for using chemicals, and efforts are needed to produce cellulase enzymes economically through media optimization. This research aims to find alternative media that make it possible to produce cellulase enzymes with high activity. This research created a control media (M0) using pro-analysis chemicals and CMC as a carbon source. Three types of cellulolytic bacterial growth media were also created, consisting of technical chemicals, rice straw as a substrate, and different protein sources. M1: alternative media using urea as a protein source; M2: alternative media using powdered milk as a protein source; and M3: alternative media using chicken liver as a protein source. A total of 1% of cellulolytic bacteria (Actinobacillus sp.) was grown in each media and then incubated at 40°C for 33 hours at a speed of 140 rpm, and the production curve and cellulase enzyme activity were measured at every 3-hour interval. The results showed that the highest cellulase enzyme production was achieved at 24 hours using alternative media with powdered milk (M2) as a protein source. The cellulase enzyme activity produced was 2.9612 µ/ml.

A Study on The Capabilities of Cellulolytic Bacteria in Degrading Feed Fiber

The utilization of local feedstuffs is a cost-effective strategy to reduce production expenses in animal husbandry. Enhancing the nutritional quality of these feedstuffs through fermentation presents a viable approach. This study investigates the efficacy of cellulolytic bacteria in degrading fiber in three local feed materials: duckweed (Lemna minor), water hyacinth (Eichornia crassipes), and lamtoro (Leucaena leucocephala). We employed a completely randomized block design with feed types as the block and levels of starter culture as treatment. Pediococcus pentosaceus, identified for its cellulase enzyme production with a clear zone of 4 cm on starch media, was utilized as the starter culture. The experimental procedure involved spraying Pediococcus pentosaceus onto duckweed, water hyacinth, and lamtoro, followed by incubation. The results indicated that Pediococcus pentosaceus effectively produced cellulolytic enzymes and significantly reduced crude fiber in duckweed by 18.18%, in water hyacinth by 9.17%, and in lamtoro by 4.23% (p < 0.05). Additionally, the crude protein content increased in all samples: by 4.69% in duckweed, 2.71% in water hyacinth, and 2.13% in lamtoro (p < 0.05). Among the treatments, the optimal outcome was achieved with 3 ml of Pediococcus pentosaceus. This study demonstrates the potential of Pediococcus pentosaceus for improving the nutritional profile of local feedstuffs through fermentation.

Statistical Optimization and Purification of Cellulase Enzyme Production from Trichosporon insectorum

Enzymatic degradation of cellulosic biomass represents the most sustainable and environmentally friendly method for producing liquid biofuel, widely utilized in various commercial processes. While cellulases are predominantly produced by bacteria and fungi, the enzymatic potential of cellulase-producing yeasts remains significantly less explored. In this study, the yeast strain Trichosporon insectorum, isolated from the gut of the coprophagous beetle Gymnopleurus sturmii, was utilized for cellulase production in submerged fermentation. A central composite design was employed to optimize cellulase production, with substrate concentration, temperature, and pH as dependent variables. The highest CMCase activity of 0.71 IU/mL was obtained at 1% substrate concentration, pH 5, and an incubation temperature of 40 °C for 72 h of fermentation using cellulose as a carbon source. For FPase production, the high value was 0.23 IU/mL at 0.5% CMC, pH 6, and an incubation temperature of 40 °C for 72 h. After purification, the enzymes produced by T. insectorum represent 39% of the total proteins. The results of this study offer an alternative strategy for utilizing various carbon sources, both soluble (CMC, carboxymethylcellulose) and insoluble (cellulose), to efficiently produce cellulase for the degradation of lignocellulosic materials. This approach holds promising benefits for sustainable waste management.

Isolation and Identification of Swan (Cygnus olor) Digestive Tract Cellulothic Bacteria to Support Fiber Degradation

Cellulolytic bacteria are bacteria that produce cellulase enzymes capable of degrading cellulose substrates. This study aimed to isolate and identify as well as measure the cellulase enzyme activity of cellulolytic bacteria isolated from the digestive tract of swans (Cygnus olor) in Mataram. The bacteria were isolated using de Man Rogosa Sharpe (MRS) selective media with the spread plate method. Then, the extracellular enzyme activity test was conducted by growing selected pure isolates on 1% Carboxy Methyl Cellulose (CMC 1%) media, followed by pouring 0.1% congo red solution and rinsing with 1M NaCl solution to determine cellulolytic activity (cellulolytic potential indicated by the appearance of a clear zone around the colony) and measuring the cellulolytic index. The bacterial isolation results obtained 4 isolates with the potential to support fiber degradation in animal feed. The highest cellulolytic index was produced by the isolate coded S1, reaching 34.5 mm, while the lowest cellulolytic index was produced by 2 isolates coded J21 and I2, reaching 18.8 mm.

Extremozymes and compatible solute production potential of halophilic and halotolerant bacteria isolated from crop rhizospheric soils of Southwest Saurashtra Gujarat

Halophiles are one of the classes of extremophilic microorganisms that can flourish in environments with very high salt concentrations. In this study, fifteen bacterial strains isolated from various crop rhizospheric soils of agricultural fields along the Southwest coastline of Saurashtra, Gujarat, and identified by 16S rRNA gene sequencing as Halomonas pacifica, H. stenophila, H. salifodinae, H. binhaiensis, Oceanobacillus oncorhynchi, and Bacillus paralicheniformis were investigated for their potentiality to produce extremozymes and compatible solute. The isolates showed the production of halophilic protease, cellulase, and chitinase enzymes ranging from 6.90 to 35.38, 0.004–0.042, and 0.097–0.550 U ml−1, respectively. The production of ectoine-compatible solute ranged from 0.01 to 3.17 mg l−1. Furthermore, the investigation of the ectoine-compatible solute production at the molecular level by PCR showed the presence of the ectoine synthase gene responsible for its biosynthesis in the isolates. Besides, it also showed the presence of glycine betaine biosynthetic gene betaine aldehyde dehydrogenase in the isolates. The compatible solute production by these isolates may be linked to their ability to produce extremozymes under saline conditions, which could protect them from salt-induced denaturation, potentially enhancing their stability and activity. This correlation warrants further investigation.

Optimization, gene cloning, expression, and molecular docking insights for enhanced cellulase enzyme production by Bacillus amyloliquefaciens strain elh1

BackgroundIn this study, we isolated a cellulase-producing bacterium, Bacillus amyloliquefaciens strain elh, from rice peel. We employed two optimization methods to enhance the yield of cellulase. Firstly, we utilized a one-variable-at-a-time (OVAT) approach to evaluate the impact of individual physical and chemical parameters. Subsequently, we employed response surface methodology (RSM) to investigate the interactions among these factors. We heterologously expressed the cellulase encoding gene using a cloning vectorin E. coli DH5α. Moreover, we conducted in silico molecular docking analysis to analyze the interaction between cellulase and carboxymethyl cellulose as a substrate.ResultsThe bacterial isolate eh1 exhibited an initial cellulase activity of 0.141 ± 0.077 U/ml when cultured in a specific medium, namely Basic Liquid Media (BLM), with rice peel as a substrate. This strain was identified as Bacillus amyloliquefaciens strain elh1 through 16S rRNA sequencing, assigned the accession number OR920278 in GenBank. The optimal incubation time was found to be 72 h of fermentation. Urea was identified as the most suitable nitrogen source, and dextrose as the optimal sugar, resulting in a production increase to 5.04 ± 0.120 U/ml. The peak activity of cellulase reached 14.04 ± 0.42 U/ml utilizing statistical optimization using Response Surface Methodology (RSM). This process comprised an initial screening utilizing the Plackett–Burman design and further refinement employing the BOX -Behnken Design. The gene responsible for cellulase production, egl, was effectively cloned and expressed in E. coli DH5α. The transformed cells exhibited a cellulase activity of 22.3 ± 0.24 U/ml. The egl gene sequence was deposited in GenBank with the accession number PP194445. In silico molecular docking revealed that the two hydroxyl groups of carboxymethyl cellulose bind to the residues of Glu169 inside the binding pocket of the CMCase. This interaction forms two hydrogen bonds, with an affinity score of −5.71.ConclusionsOptimization of cultural conditions significantly enhances the yield of cellulase enzyme when compared to unoptimized culturing conditions. Additionally, heterologous expression of egl gene showed that the recombinant form of the cellulase is active and that a valid expression system can contribute to a better yield of the enzyme.

Isolation and Characterization of Aeromonas taiwanensis Strain for Simultaneous Production of Cellulase, Amylase, Pectinase, and Protease Enzymes

Isolation and Characterization of Aeromonas taiwanensis Strain for Simultaneous Production of Cellulase, Amylase, Pectinase, and Protease Enzymes

Optimization of Fermentation Conditions for Cellulase Production by Trichoderma harzianum PK5 Obtained from Decaying Palm Kernel Cake

Study’s Novelty/ Excerpt This study successfully optimized cellulase production by Trichoderma harzianum PK5 using the One Factor at a Time (OFAT) approach, identifying copra meal and KNO3 as the best carbon and nitrogen sources. Optimal conditions included a pH of 4.0, 125% moisture concentration, 8% inoculum size, 30°C temperature, and a 7-day incubation period, yielding a high enzyme titre of 252.54±7.73 U/gds in solid-state fermentation. These findings highlight the potential of T. harzianum PK5 as a cost-effective source of cellulases for industrial applications. Full Abstract Cellulases are considered to be among the most important enzymes in the commercial market and in various industries. Their applications are widespread, leading to increased demand and high associated costs. This necessitates the search for more cost-effective cellulases from microorganisms. Therefore, the aim of this study was to optimize cellulase production by Trichoderma harzianum PK5 using the One Factor at a Time (OFAT) approach. The effects of carbon, nitrogen, and various environmental factors were studied in both submerged and solid-state fermentation setups by adjusting one factor at a time based on the optimal conditions established from the previous condition. Copra meal and KNO3 were identified as the best complex carbon and nitrogen sources, respectively, for cellulase production by Trichoderma harzianum PK5. The optimal pH of 4.0, moisture concentration of 125% (v/w), inoculum size of 8%, temperature of 30°C, and an incubation time of 7 days were determined as the optimal conditions for cellulase production by this isolate, resulting in an enzyme titre of 252.54±7.73 U/gds in solid-state fermentation. It was found that cellulase enzyme production by the isolate was constitutive. In conclusion, cellulase production by T. harzianum PK5 was significantly optimized using the OFAT approach

Extracellular Endoglucanase and Exoglucanase Enzymes Production by Trichoderma viride Utilizing Olive Mill Wastewater (OMW) in liquid fermentation

Background:The cellulase enzyme is one of the most industrially important enzymes. Its cost represents a significant barrier to other valuable commercial products. Reducing the cost of cellulase production is an important approach. Objectives: For this purpose, this work investigated the production of cellulase enzyme using olive oil production waste (olive mill wastewater (OMW). Materials and methods: The ability of Trichoderma viride to utilize OMW as a substrate for cellulase production was studied. Optimization of cellulase production was investigated to find out the optimum OMW concentration, agitation speed, aeration rate, and cellulose addition. Results:The results showed that 75% v/v OMW submerged shake flask culture was the most suitable culture for T. viride growth and cellulase enzyme production (0.82 U/ml endoglucanase activity and 0.25 U/ml exoglucanase activity). When this culture was supplemented with cellulose, the activity of endoglucanase and exoglucanase was significantly improved (10.24 U/ml and 2.17 U/ml respectively).The agitation speed of 200 rpm enhanced the production to reach 9.1 U/ml of endoglucanase and 6.38 U/ml of exoglucanase. The effect of the aeration rate on enzyme production was studied under batch cultivation. The highest cellulase activity was at 2.0 vvm, where the endoglucanase and exoglucanase activities were 55.96 U/ml and 32.62 U/ml respectively. Conclusions: Therefore, it is claimed that OMW is a suitable medium for cellulase enzyme production after optimization of the process.

Production of cellulase enzymes by <i>Bacillus</i> sp and <i>Pseudomonas</i> sp isolated from anthill soil

Cellulase turns the most widespread biopolymer and biologically sustainable resource, 'cellulose,' into reducing sugar. The study aimed at producing cellulase enzymes by bacteria isolated from anthill soil. Cellulase-producing bacteria were isolated from anthill soil using Carboxymethyl cellulose (CMC) medium. The isolates were screened for cellulase production by cultural, morphological, biochemical and sugar fermentation tests. Optimization of the fermentation medium for maximum cellulase production was carried out by one factor at a time (OFAT). Data obtained were analysed with the analysis of variance (ANOVA) using SPSS 2007, version 16.0. The identified Pseudomonas sp, Staphylococcus sp, E. coli and Bacillus sp were isolated with highest potential of cellulase production. The culture conditions like pH, temperature, carbon sources and nitrogen sources were optimized. The optimum conditions found for cellulase production were 40oC at pH 8.5 with maltose as carbon source and yeast extract as nitrogen source. The highest activity and stability of cellulase enzymes between neutral to alkaline pH and high temperature.

Green ecofriendly enhancement of cellulase productivity using agricultural wastes by Aspergillus terreus MN901491: statistical designs and detergent ability on cotton fabrics

BackgroundCellulase is considered a group member of the hydrolytic enzymes, responsible for catalyzing the hydrolysis of cellulose and has various industrial applications. Agricultural wastes are used as an inexpensive source for several utilizable products throughout the world. So, searching for cellulase enzymes from fungal strains capable of utilizing agricultural wastes to increase productivity, reduce costs and overcome waste accumulation in the environment is very important to evaluate its potency as a bio-additive to detergent agents.ResultsIn the current study, the previously identified fungal strain Aspergillus terreus MN901491 was screened and selected for cellulase production. Medium parameters were optimized using one-factor-at-a-time (OFAT) and multi-factorial (Plackett-Burman and Box-Behnken) design methods. OFAT showed the ability of the fungal strain to utilize agricultural wastes (corn cob and rice straw) as a substrate. Also, yeast extract was the best nitrogen source for enhancing cellulase productivity. The most significant variables were determined by Plackett-Burman Design (PBD) and their concentrations were optimized by Response Surface Methodology (RSM) using Box-Behnken Design (BBD). Among eleven independent variables screened by PBD, malt extract, (NH4)2SO4, and KCl were the most significant ones followed by rice straw which affected cellulase production positively. The ANOVA results particularly the R2-value of PBD (0.9879) and BBD (0.9883) confirmed the model efficiency and provided a good interpretation of the experiments. PBD and BBD improved cellulase productivity by 6.1-fold greater than that obtained from OFAT. Medium optimization using OFAT and statistical models increased cellulase production from A. terreus MN901491 by 9.3-fold compared to the non-optimized medium. Moreover, the efficiency of cellulase activity on cotton fabrics as a bio-additive detergent was evaluated and estimated using whiteness and scanning electron microscope (SEM) that affirmed its potential effect and remarkable detergent ability to improve whiteness by 200% in comparison with non-washed fabric and by 190% in comparison with fabric washed by water.ConclusionThe presented work was stabilized as a multi-efficiency in which wastes were used to produce cellulase enzyme from the fungal strain, Aspergillus terreus MN901491 as a bio-additive to detergent applications that involved ecofriendly and green processes.

Surface-Enhanced Raman Spectroscopy for Monitoring the Biochemical Changes Due to DNA Mutations Induced by CRISPR-Cas9 Genome Editing in the Aspergillus niger Fungus.

In this study, surface-enhanced Raman spectroscopy (SERS) technique, along with principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA), is used as a simple, quick, and cost-effective analysis method for identifying biochemical changes occurring due to induced mutations in the Aspergillus niger fungus strain. The goal of this study is to identify the biochemical changes in the mutated fungal cells (cell mass) as compared to the control/nonmutated cells. Furthermore, multivariate data analysis tools, including PCA and PLS-DA, are used to further confirm the differentiating SERS spectral features among fungal samples. The mutations are caused in A. niger by the clustered regularly interspaced palindromic repeat CRISPR-Cas9 genomic editing method to improve their biotechnological potential for the production of cellulase enzyme. SERS was employed to detect the changes in the cells of mutated A. niger fungal strains, including one mutant producing low levels of an enzyme and another mutant producing high levels of the enzyme as a result of mutation as compared with an unmutated fungal strain as a control sample. The distinctive features of SERS corresponding to nucleic acids and proteins appear at 546, 622, 655, 738, 802, 835, 959, 1025, 1157, 1245, 1331, 1398, and 1469 cm-1. Furthermore, PLS-DA is used to confirm the 89% accuracy, 87.7% precision, 87% sensitivity, and 88.9% specificity of this method, and the value of the area under the curve (AUROC) is 0.67. It has been shown that surface-enhanced Raman spectroscopy is an effective method for identifying and differentiating biochemical changes in genome-modified fungal samples.

Development and evaluation of agro-waste biodegradation formulation

In the study, twenty strains of cellulose degrading microbes (Pseudomonas spp., Bacillus spp., Aspergillus spp. etc.) were isolated from the gut of termites and earthworms collected from different locations. The cellulolytic activity of the isolates of Aspergillus spp. were determined based on the formation of clear zone on CMC (Carboxymethyl cellulose) media. The strain C18 of Aspergillus flavus isolated from the gut of termite produced the largest zone with maximum cellulolytic index of 7.64. All the purified strains were subjected to in vitro cellulase activity through DNS (3,5-dinitrosalicylic acid) method which showed that strain C18 of Aspergillus flavus produced the maximum amount of glucose (0.507 mg ML ) and the -1 cellulase enzyme ( IU ML ) was recorded as 0.187 IU ML . Out of twenty microbial strains, ten best -1 -1 microbial strains were selected and subjected to in vitro FPA (filter paper assay). Aspergillus flavus strain C18 was found most effective with maximum glucose (1.059 mg ml ) and cellulase enzyme (0.196 -1 IUML ) production after 9 days of inoculation and therefore the consortium was made from two most -1 effective Aspergillus flavus strains (C12 and C18) which were found best effective in degradation of wheat straw substrate i.e., 61 per cent degradation of initial substrate weight. After evaluation of each strain, effective strains (C12 and C18) were used to prepare consortium formulations for commercial use in the field. These cellulolytic microbes have significant role in agro-waste management by degradation without harming the land and environment.. KEYWORDS :Agro-waste, Aspergillus flavus, biodegradation, cellulolytic microbes, formulation

PH and Temperature Optimization of Several Bacterial Isolates from Mangrove Waters in the Mandeh Area to Produce Cellulase Enzyme

pH and Temperature Optimization of Several Bacterial Isolates from Mangrove Waters in the Mandeh Area to Produce Cellulase Enzyme

Characterization of Thermostable Cellulase Enzyme Isolated from a Hot Spring Bacterium: Bacillus sp.

Cellulase is a complex of enzymes which consists of β-1,4-endoglucanase, cellobiohydrolase, and β-glucosidase. Cellulase contributes a significant share to the world enzyme market and is used in number of industries viz; paper and pulp, food and beverage, bioethanol, detergent and textile. The harsh industrial conditions such as high temperature, extreme pH levels and high substrate concentration etc. which is used in such industries adversely effect on structure and activity of enzymes. Therefore, huge amount of chemical catalysts which cause chemical wastes are used in industrial settings instead of enzymes. Since the chemical wastes adversely affect the ecosystem, isolation and characterization of thermostable cellulase enzyme-producing bacteria and exploring the industrial perspective of thermostable enzymes is a better approach towards green chemistry. The present study focused on characterization of cellulase enzyme produced by Bacillus sp., which shows optimum activity at 60 C under neutral conditions, isolated from Gomarankadavala hot spring in Sri Lanka. The effect of different Carbon sources: glucose and lactose, Nitrogen sources: peptone, tryptone, yeast extract and urea on enzyme production and different Carboxymethylcellulose (CMC) concentrations; 0.5%, 1.5% and 2%, on enzyme activity were measured using the DNS method. Under the optimum conditions, cellulase enzyme activity on different substrates: corn cob, rice bran and raw leaves were measured. The highest enzyme production was recorded in the culture medium which added tryptone as the nitrogen source and adding carbon sources to the culture medium showed an increase of cellulase enzyme production. The optimum CMC concentration for the enzyme activity was recorded as 1% and from all the optimized parameters, the substrate; raw leaves showed the highest enzyme activity of 11.305 U/ml. Further, a considerable amount of enzyme activity was recorded on corn cobs and rice bran as well. Thus, the thermostable cellulase enzyme produced by bacterium identified as Bacillus cereus isolated from Gomarankadavala hot spring could be successfully used in industrial settings such as kitchen/industrial waste management, bioethanol production, paper and pulp industry and textile industry which use high temperatures and cellulosic substrates as raw materials. 
 Keywords: Thermostable enzymes, Cellulase, Industrial applications, Hot springs

Marine cyanobacterial biomass is an efficient feedstock for fungal bioprocesses

BackgroundMarine cyanobacteria offer many sustainability advantages, such as the ability to fix atmospheric CO2, very fast growth and no dependence on freshwater for culture. Cyanobacterial biomass is a rich source of sugars and proteins, two essential nutrients for culturing any heterotroph. However, no previous study has evaluated their application as a feedstock for fungal bioprocesses.ResultsIn this work, we cultured the marine cyanobacterium Synechococcus sp. PCC 7002 in a 3-L externally illuminated bioreactor with working volume of 2 L with a biomass productivity of ~ 0.8 g L−1 day−1. Hydrolysis of the biomass with acids released proteins and hydrolyzed glycogen while hydrolysis of the biomass with base released only proteins but did not hydrolyze glycogen. Among the different acids tested, treatment with HNO3 led to the highest release of proteins and glucose. Cyanobacterial biomass hydrolysate (CBH) prepared in HNO3 was used as a medium to produce cellulase enzyme by the Penicillium funiculosum OAO3 strain while CBH prepared in HCl and treated with charcoal was used as a medium for citric acid by Aspergillus tubingensis. Approximately 50% higher titers of both products were obtained compared to traditional media.ConclusionsThese results show that the hydrolysate of marine cyanobacteria is an effective source of nutrients/proteins for fungal bioprocesses.

In vivo application of potent probiotics for enhancing potato growth and controlling Ralstonia solanacearum and Fusarium oxysporum infections

Plant probiotics are live microbial cells or cultures that support plant growth and control plant pathogens through different mechanisms. They have various effects on plants, including plant growth promotion through the production of indole acetic acid (IAA), biological control activity (BCA), and production of cellulase enzymes, thus inducing systemic resistance and increasing the availability of mineral elements. The present work aimed to study the potential of Achromobacter marplatensis and Bacillus velezensis as plant probiotics for the field cultivation of potatoes. In vitro studies have demonstrated the ability of selected probiotics to produce IAA and cellulase, as well as antimicrobial activity against two plant pathogens that infect Solanum tuberosum as Fusarium oxysporum and Ralstonia solanacearum under different conditions at a broad range of different temperatures and pH values. In vivo study of the effects of the probiotics A. marplatensis and B. velezensis on S. tuberosum plants grown in sandy clay loamy soil was detected after cultivation for 90 days. Probiotic isolates A. marplatensis and B. velezensis were able to tolerate ultraviolet radiation (UV) exposure for up to two hours, the dose response curve exhibited that the D10 values of A. marplatensis and B. velezensis were 28 and 16 respectively. In the case of loading both probiotics with broth, the shoot dry weight was increased significantly from 28 in the control to 50 g, shoot length increased from 24 to 45.7 cm, branches numbers increased from 40 to 70 branch, leaves number increased from 99 to 130 leaf, root dry weight increased from 9.3 to 12.9 g, root length increased from 24 to 35.7 cm, tuber weight increased from 15 to 37.0 g and tubers number increased from 9 to 24.4 tuber, the rot percentage was reduced to 0%. The addition of both probiotic isolates, either broth or wheat grains load separately has enhanced all the growth parameters; however, better results and increased production were in favor of adding probiotics with broth more than wheat. On the other hand, both probiotics showed a remarkable protective effect against potato pathogens separately and reduced the negative impact of the infection using them together.

Isolation and Characterization of Lactic Acid Bacteria from Rice Washing Water Waste

Rice washing water is still considered as household waste because its benefits for plants are not widely known. Rice washing water is obtained from water that is washed many times before cooking which is rich in carbohydrate content in the form of starch and glucose and vitamins. Lactic acid bacteria (LAB) require sugar or carbohydrates from rice washing water which is used as food to support metabolic activity and microbial cell proliferation. LAB is also included in a large group of microorganisms that physiologically produce lactic acid as the main metabolic product. LAB are classified as good bacteria so they are widely used as food processing products, especially those related to human digestion. The use of lactic acid bacteria in the agricultural sector is still very minimal, especially as a biodecomposer, even though LAB has the ability to produce cellulase enzymes which are very important in the breakdown of lignin in organic matter. This study aims to determine the presence of lactic acid bacteria in rice washing water fermentation. Isolation of lactic acid bacteria using pour plate method with MRSA (de Man Rogosa Sharpe Agar) media. Lactic acid bacteria isolates obtained were then characterized morphologically, physically, and biochemically. The results of isolation and characterization obtained four isolates with the codes AL-01, AL-02, AL-03, and AL-04. Four LAB isolates showed gram-positive, catalase-positive, non-motile, non-H2S, and IAA-positive results.

Screening and characterization of endophytic bacteria isolated from celery with the potential to promote plant growth

Abstract. Nuriyanah, Widowati T, Masnang A, Nurjanah L, Novilasari D, Lekatompessy SJR, Simarmata R. 2023. Screening and characterization of endophytic bacteria isolated from celery with the potential to promote plant growth. Biodiversitas 24: 6897-6904. Endophytic bacteria that inhabit plant tissue contribute to the enhancement of host plant growth by producing secondary metabolites. They can improve plant growth by phytohormone modulation and nutrient uptake. In addition, endophytic bacteria can increase plant health and tolerance by antibiotic and hydrolytic enzyme production. This study aims to screen and characterize endophytic bacteria of celery as plant growth promoter agents. Thirty endophytic bacteria were isolated successfully from part of the celery plant using the spread plate and plant piece method which grew at Nutrient Agar media. Based on the characterization assay, 30 isolates may produce IAA with various concentrations of 0.21-7.18 mg/L. In addition, 3 of 30 isolates had phosphate solubilizing activity, in different indexes ranging from 0.71-1.43. Twenty-one isolates were able to grow in an N-free medium, indicating the capability of isolates to fix nitrogen qualitatively. The hydrolytic enzymes assay resulted in 14 amylolytic, 17 proteolytic, and 12 cellulolytic activities. In addition, nine of 30 isolates show multi-activity in amylase, protease, and cellulase enzyme production. Based on the screening results, there are four potential isolates (SLBg 1.2; SLBg 1.5; SLAg 1.1, and SLAg 1.5) as promising plant-growth promoters and were molecularly identified as members of the Bacillus genus. These isolates can be developed as biofertilizers for supporting the cultivation of celery plants.