scholarly journals Optimization of Cellulase Production by a Novel Endophytic Fungus Penicillium oxalicum R4 Isolated from Taxus cuspidata

2021 ◽  
Vol 13 (11) ◽  
pp. 6006
Author(s):  
Hongkun Li ◽  
Meijia Dou ◽  
Xinyu Wang ◽  
Na Guo ◽  
Ping Kou ◽  
...  

Endophytic fungi inside a plant can degrade a portion of plant lignin and cellulose. Endophytic Penicillium is one of the industrial microorganisms with the advantage of producing enzymes with a complete enzyme system that can be secreted into the extracellular space. The natural evolution of ancient tree species from special natural geographic environments to screen out cellulase-producing strains with excellent characteristics provides a promising direction for future industrial enzymes. The present study successfully isolated and screened a novel fungal endophyte, Penicillium oxalicum R4, with higher cellulase activity from Taxus cuspidata. Under the optimized culture conditions obtained by a Box–Behnken design (BBD) and an artificial neural network–genetic algorithm (ANN–GA), yields of Filter Paperase (FPase), Carboxymethyl Cellulase (CMCase) and β-glucosidase (βGLase) produced by P. oxalicum R4 were 1.45, 5.27 and 6.35 U/mL, which were approximately 1.60-fold, 1.59-fold and 2.16-fold higher than those of the non-optimized culture, respectively. The discovery of cellulase-producing strains of endophytic fungi located in special natural geographic environments, such as Taxus cuspidata, which is known as a living plant fossil, provides new research directions for future industrial enzymes.

2011 ◽  
Vol 183-185 ◽  
pp. 994-998
Author(s):  
Shuo Dong ◽  
Nai Yu Chi ◽  
Qing Fang Zhang

The design of an optimum and cost-efficient medium for production of cold-active cellulase by Penicillium cordubense D28 was attempted by using response surface methodology (RSM). Based on the Plackett–Burman design, corn meal, (NH4)2SO4 and branc were selected as the most critical nutrient. Subsequently, they were investigated by the Box-Behnken design. Results showed that the maximum cold-active cellulase activity of 110.4U/mL was predicted when the concentration of corn meal, (NH4)2SO4 and branc were 21.97 g/L, 2.39 g/L and 14.99 g/L, respectively. The results were further verified by triplicate experiments. The batch reactors were operated under an optimized condition of the respective corn meal, (NH4)2SO4 and branc concentration of 22 g/L , 2.4 g/L and 15 g/L , the initial pH of 6.0 and experimental temperature of 20 ± 1°C. Without further pH adjustment, the maximum cold-active cellulase activity of 109.8 U/mL was obtained based on the optimized medium with further verified the practicability of this optimum strategy.


2021 ◽  
Vol 4 (3) ◽  
pp. 1-3
Author(s):  
Sohail Khan ◽  
Ashwani Mathur

The growing demand and application of industrially important enzyme necessitate the need to explore new sources with diverse enzymes ranging in their specificity and activities. Enzymes are safe alternatives to chemical synthesis due to minimum side effect and ease of manufacturing. Solid state fermentation (SSF) is a cost-effective alternative to submerged fermentation with agro-residues or waste, often being used as substrate for growing diverse organisms for production of metabolites. Current study is one of the scarce report on exploring alpha amylase and cellulase production ability Aspergillus ochraceus (MTCC 1877) using wheat bran as substrate at relative humidity of 90% and at 30 ºC, for 7 days. Result showed the potential of Aspergillus ochraceus (MTCC 1877), as potential source of the two enzymes. Results revealed comparatively higher alpha amylase activity in the SSF extract of Aspergillus ochraceus (MTCC 1877) in comparison to Trichoderma longibrachiatum (ITCC 7839). On the contrary, comparatively higher cellulase activity was observed in the SSF extract of Trichoderma longibrachiatum (ITCC 7839). The results showed the potential of Aspergillus ochraceus (MTCC 1877) as a source of the two enzymes. Variation in enzymes activity may be attributed to the experimental culture conditions and may be further optimized to enhance the enzymes yield.


2012 ◽  
Vol 518-523 ◽  
pp. 5578-5585
Author(s):  
Nan Ban ◽  
Yu Jie Zhou ◽  
Yan Ping Ye ◽  
Lin Mei Dai ◽  
Alatangaole Damirn ◽  
...  

Plackett-Burman design was employed for screening culture conditions for cellulase production by Penicillium decumbens in submerged fermentation. The results showed that wheat bran was the most significant factor influencing Filter Paper Activity (FPA) of the cellulase, followed by cellulose microcrystalline and initial pH, which could be further optimized for improving the cellulase activity. The effects of pH and temperature on FPA assay were investigated, and optimal FPA could be obtained at pH 4.5 and 60 °C. The stabilities of endo-glucanase (EG), exo-glucanase (CBH) and β-glucosidase (BG) were investigated and compared with that of FPA under different pH and temperature. The results indicated that CBH and FPA were more sensitive to pH and temperature than EG and BG and the stability of CBH was very similar to that of FPA under the conditions.


2000 ◽  
Vol 26 (5-6) ◽  
pp. 394-401 ◽  
Author(s):  
F.C. Domingues ◽  
J.A. Queiroz ◽  
J.M.S. Cabral ◽  
L.P. Fonseca

Phytotaxa ◽  
2021 ◽  
Vol 514 (3) ◽  
pp. 247-260
Author(s):  
KASUN THAMBUGALA ◽  
DINUSHANI DARANAGAMA ◽  
SAGARIKA KANNANGARA ◽  
THENUKA KODITUWAKKU

Endophytic fungi are a diverse group of microorganisms that live asymptomatically in healthy tissues of host and they have been reported from all kinds of plant tissues such as leaves, stems, roots, flowers, and fruits. In this study, fungal endophytes associated with tea leaves (Camellia sinensis) were collected from Kandy, Kegalle, and Nuwara Eliya districts in Sri Lanka and were isolated, characterized, and identified. A total of twenty endophytic fungal isolates belonging to five genera were recovered and ITS-rDNA sequence data were used to identify them. All isolated endophytic fungal strains belong to the phylum Ascomycota and the majority of these isolates were identified as Colletotrichum species. Phyllosticta capitalensis was the most commonly found fungal endophyte in tea leaves and was recorded in all three districts where the samples were collected. This is the very first investigation on fungal endophytes associated with C. sinensis in Sri Lanka based on molecular sequence data. In addition, a comprehensive account of known endophytic fungi reported worldwide on Camellia sinensis is provided.


2021 ◽  
Author(s):  
Oladipo Olaniyi

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.


2020 ◽  
Vol 36 (4) ◽  
Author(s):  
Ega Soujanya Lakshmi ◽  
Manda Rama Narasinga Rao ◽  
Muddada Sudhamani

ABSTRACT Thirty seven different colonies were isolated from decomposing logs of textile industries. From among these, a thermotolerant, grampositive, filamentous soil bacteria Streptomyces durhamensis vs15 was selected and screened for cellulase production. The strain showed clear zone formation on CMC agar plate after Gram’s iodine staining.  Streptomyces durhamensis vs15 was further confirmed for cellulase production by estimating the reducing sugars through dinitrosalicylic acid (DNS) method. The activity was enhanced by sequential mutagenesis using three mutagens of ultraviolet irradiation (UV), N methyl-N’-nitro-N-nitrosoguanidine (NTG) and Ethyl methane sulphonate (EMS). After mutagenesis, the cellulase activity of GC23 (mutant) was improved to 1.86 fold compared to the wild strain (vs15). Optimal conditions for the production of cellulase by the GC 23 strain were evaluated using Response Surface Methodology (RSM) and Artificial Neural Network (ANN). Effect of pH, temperature, duration of incubation, , and substrate concentration on cellulase production were evaluated. Optimal conditions for the production of cellulase enzyme using Carboxy Methyl Cellulase as a substrate are 55 oC of temperature, pH of 5.0 and incubation for 40 h. The cellulase activity of the mutant Streptomyces durhamensis GC23 was further optimised to 2 fold of the activity of the wild type by RSM and ANN.  


mBio ◽  
2020 ◽  
Vol 11 (6) ◽  
Author(s):  
Logan A. Collier ◽  
Arit Ghosh ◽  
Katherine A. Borkovich

ABSTRACT The filamentous fungus Neurospora crassa decomposes lignocellulosic biomass to generate soluble sugars as carbon sources. In this study, we investigated a role for heterotrimeric G-protein signaling in cellulose degradation. Loss of the Gα subunit genes gna-1 and gna-3, the Gβ subunit genes gnb-1 and cpc-2, the Gγ gene gng-1, or the gene for downstream effector adenylyl cyclase (cr-1) resulted in loss of detectable cellulase activity. This defect was also observed in strains expressing a constitutively active version of gna-3 (gna-3Q208L). We found that GNA-1 levels are greatly reduced in Δgna-3, Δgnb-1, and Δgng-1 strains, likely contributing to cellulase defects in these genetic backgrounds. The observation that gna-3Q208L Δgnb-1 strains exhibit cellulase activity, despite greatly reduced levels of GNA-1 protein, is consistent with positive control of cellulase production by GNA-3 that is manifested in the absence of gnb-1. Expression patterns for five cellulase genes showed that Δgna-1, Δgnb-1, and Δgna-3 mutants produce less cellulase mRNA than the wild type, consistent with transcriptional regulation. Δcpc-2 mutants had wild-type levels of cellulase transcripts, suggesting posttranscriptional control. In contrast, results for Δcr-1 mutants support both transcriptional and posttranscriptional control of cellulase activity by cAMP signaling. Cellulase activity defects in Δgna-3 mutants were fully remediated by cAMP supplementation, consistent with GNA-3 operating upstream of cAMP signaling. In contrast, cAMP addition only partially corrected cellulase activity defects in Δgna-1 and Δgnb-1 mutants, suggesting participation of GNA-1 and GNB-1 in additional cAMP-independent pathways that control cellulase activity. IMPORTANCE Filamentous fungi are critical for the recycling of plant litter in the biosphere by degrading lignocellulosic biomass into simpler compounds for metabolism. Both saprophytic and pathogenic fungi utilize plant cell wall-degrading enzymes to liberate carbon for metabolism. Several studies have demonstrated a role for cellulase enzymes during infection of economically relevant crops by fungal pathogens. Especially in developing countries, severe plant disease means loss of entire crops, sometimes leading to starvation. In this study, we demonstrate that G-protein signaling is a key component of cellulase production. Therefore, understanding the role of G-protein signaling in the regulation of the unique metabolism of cellulose by these organisms can inform innovations in strain engineering of industrially relevant species for biofuel production and in combatting food shortages caused by plant pathogens.


2020 ◽  
Vol 20 ◽  
pp. 03005
Author(s):  
Safinah Surya Hakim ◽  
Tri W. Yuwati

Penicillium citrinum is an endophyte fungus isolated from plant tissues that live in tropical peatland. Various study reveals that endophytic fungi give advantages to plant health. Since 2015, series of experiments were conducted to investigate the capability of P. citrinum P3.10 as biofertilizer of plant tree in nursery stage which are: pathogenicity test, in vitro assay, direct application to seeds, dosage test, and fertilizer formulation. This paper presents a review from our research of P. citrinum isolate P3.10 from the isolation stage to the biofertilizer product formulation. Resear ch results showed that the application of endophytic fungi to seedlings gave positive responses. However, during the research process there were also obstacles in the application of endophytic fungi to plants, which are: (a) difficulties in application due to the varied response, (b) formulation, (c) storage, (d) isolate viability, and (d) fungal isolate maintenance. Risk management was needed during biofertilizer production. Hence, the utilization of biofertilizer made from endophyte fungi is applicable and can be used not only on an experimental scale but also on a larger scale.


2018 ◽  
Vol 67 ◽  
pp. 02051
Author(s):  
Misri Gozan ◽  
Andre Fahriz Harahap ◽  
Chandra Paska Bakti ◽  
Siswa Setyahadi

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.


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