Cassava starch is considered as a potential source for the commercial production of bioethanol because of its availability and low market price. It can be used as a basic source to support large-scale biological production of bioethanol using microbial amylases. With the progression and advancement in enzymology, starch liquefying and saccharifying enzymes are preferred for the conversion of complex starch polymer into various valuable metabolites. These hydrolytic enzymes can selectively cleave the internal linkages of starch molecule to produce free glucose which can be utilized to produce bioethanol by microbial fermentation. In the present study, several filamentous fungi were screened for production of amylases and among them Aspergillus fumigatus KIBGE-IB33 was selected based on maximum enzyme yield. Maximum α-amylase, amyloglucosidase and glucose formation was achieved after 03 days of fermentation using cassava starch. After salt precipitation, fold purification of α-amylase and amyloglucosidase increased up to 4.1 and 4.2 times with specific activity of 9.2 kUmg-1 and 393 kUmg-1, respectively. Concentrated amylolytic enzyme mixture was incorporated in cassava starch slurry to give maximum glucose formation (40.0 gL-1), which was further fermented using Saccharomyces cerevisiae into bioethanol with 84.0% yield. The distillate originated after recovery of bioethanol gave 53.0% yield. An improved and effective dual enzymatic starch degradation method is designed for the production of bioethanol using cassava starch. The technique developed is more profitable due to its fast liquefaction and saccharification approach that was employed for the formation of glucose and ultimately resulted in higher yields of alcohol production.

/pdf/saccharification-and-liquefaction-of-cassava-starch-an-5cxp8uo8sl.pdf

Saccharification and liquefaction of cassava starch: An alternative source for the production of bioethanol using amylolytic enzymes by double fermentation process

The study aimed at isolation and screening of fungal amylase producer, optimization of solid state fermentation conditions for maximum amylase production by the best amylase producer, and characterization of the crude amylases, so produced. Aspergillus fumigatus NTCC1222 showed the highest amylase activity (164.1 U/mL) in secondary screening under SSF conditions and was selected for further studies. The test strain showed maximum amylase production (341.7 U/mL) and supernatant protein concentration (9.7 mg/mL) for incubation period (6 days), temperature (35°C), initial pH (6.0), nutrient salt solution as moistening agent, and beef extract as nitrogen source. Pomegranate peel produced maximum amylase activity, but wheat bran (only slightly lesser amylase activity as compared to that of pomegranate peel) was chosen for further studies, keeping in mind the seasonal availability of pomegranate peel. TLC confirmed the amylase produced to be α-type and 60 kDa was the molecular weight of the partially purified amylase. The enzyme showed maximum enzyme activity at pH 6.0, temperature of 55°C, and incubation time of 60 minutes. UV (616.0 U/mL) and chemical (814.2 U/mL) mutation enhanced amylase activity as compared to wild test strain. The study indicates that Aspergillus fumigatus NTCC1222 can be an important source of amylase and the crude enzyme, hence obtained, can be cost effectively applied in multiple sections of textile wet processing.

https://downloads.hindawi.com/journals/bmri/2014/215748.pdf

Production of Fungal Amylases Using Cheap, Readily Available Agriresidues, for Potential Application in Textile Industry

Production of amylase by fungi isolated from legume seeds collected in Almadinah Almunawwarah, Saudi Arabia

Environmental screening programs are used to find new enzymes that may be utilized in large-scale industrial processes. Among microbial sources of new enzymes, the rationale for screening fungal endophytes as a potential source of such enzymes relates to the hypothesised mutualistic relationship between the endophyte and its host plant. There is a need for new microbial amylases that are active at low temperature and alkaline conditions as these would find industrial applications as detergents. An α-amylase produced by Preussia minima, isolated from the Australian native plant, Eremophilia longifolia, was purified to homogeneity through fractional acetone precipitation and Sephadex G-200 gel filtration, followed by DEAE-Sepharose ion exchange chromatography. The purified α-amylase showed a molecular mass of 70 kDa which was confirmed by zymography. Temperature and pH optima were 25°C and pH 9, respectively. The enzyme was activated and stabilized mainly by the metal ions manganese and calcium. Enzyme activity was also studied using different carbon and nitrogen sources. It was observed that enzyme activity was highest (138 U/mg) with starch as the carbon source and L-asparagine as the nitrogen source. Bioreactor studies showed that enzyme activity was comparable to that obtained in shaker cultures, which encourages scale-up fermentation for enzyme production. Following in-gel digestion of the purified protein by trypsin, a 9-mer peptide was sequenced and analysed by LC-ESI-MS/MS. The partial amino acid sequence of the purified enzyme presented similarity to α-amylase from Magnaporthe oryzae. The findings of the present study indicate that the purified α-amylase exhibits a number of promising properties that make it a strong candidate for application in the detergent industry. To our knowledge, this is the first amylase isolated from a Preussia minima strain of endophytic origin.

/pdf/amylase-production-by-preussia-minima-a-fungus-of-endophytic-4ndwqp9v3d.pdf

Amylase production by Preussia minima, a fungus of endophytic origin: optimization of fermentation conditions and analysis of fungal secretome by LC-MS

Amylases are amongst the most important hydrolytic enzymes that are used in numerous industrial uses reaching for food to pharmaceuticals Immobilization of enzymes can proposal several assistances as reusability and retrieval from their products improve strength under both operating and storing environments Marine fungal isolate was recovered from red sea water at Sharm El-Sheikh Province and was tested for amylase activity using different agricultural wastes as substrate It was found that pomegranate peel was the best substrate for amylase production (339 U/ml) Thus, it was subjected for identifying by 18S rDNA gene The phylogenetic analysis results indicated that this fungal isolate belonged to Aspergillus species with similarity of 99% and named as Aspergillus terrus SS_RS-NE Its nucleotide sequences were deposited in NCBI GenBank under accession no of MN901491 Some parameters affecting amylase activity using pomegranate peel as substrate were studied The results denoted that, the highest amylase activity of 34069 U/ml using 15% pomegranate peel at 30 °C, pH 60 on 5 days incubation time by Aspergillus terreus The produced crude enzyme was partially purified with 80% ammonium sulfate followed by dialysis The enzyme activity was 1246 U/ml and 2411 U/ml employing ammonium sulfate precipitation and dialysis respectively The partially purified amylase was immobilized with 2% sodium alginate and the results showed the highest immobilized enzyme yield was 928% The characterizations of immobilized amylase were studied and the results indicated that, the maximal immobilized amylase activity was 25225 U/ml with 2% starch as a substrate at optimum pH value of 65, temperature at 60 °C and 10 min reaction time in comparison to maximal free amylase enzyme at pH 5, 50 °C after 40 min The results also indicated the immobilized amylase was stable at 60 °C for 20 min Aspergillus terrus SS_RS-NE (MN901491) was isolated and genetically identified It has the ability to produce amylase enzyme using pomegranate peel waste with a yield of 339 U/ml The crude enzyme was partially purified by ammonium sulfate followed by dialysis The maximal immobilized amylase activity of 25225 U/ml was obtained under optimized some culture conditions and medium nutrient parameters

/pdf/optimization-and-immobilization-of-amylase-produced-by-4jnhu3yoka.pdf

Optimization and immobilization of amylase produced by Aspergillus terreus using pomegranate peel waste

Five fungal isolates were screened for the production of alpha-amylase using both solid-state and submerged fermentations. The best amylase producer among them, Aspergillus niger JGI 24, was selected for enzyme production by solid-state fermentation (SSF) on wheat bran. Different carbon and nitrogen supplements were used to enhance enzyme production and maximum amount of enzyme was obtained when SSF was carried out with soluble starch and beef extract (1% each) as supplements. Further attempts to enhance enzyme production by UV induced mutagenesis were carried out. Survival rate decreased with increase in duration of UV exposure. Partial purification of the enzyme using ammonium sulphate fractionation resulted in 1.49 fold increase in the enzyme activity. The enzyme showed a molecular weight of 43 kDa by SDS-PAGE. Metal ions Ca2+ and Co2+ increased the enzyme activity. The enzyme was optimally active at 30 degrees C and pH 9.5.

Production and characterization of alpha-amylase from Aspergillus niger JGI 24 isolated in Bangalore

Background: The aerosols generated during dental treatments contain bacteria and other microorganisms that penetrate the body through the respiratory system of dental surgeons and cause infectious diseases. Several studies have been done to reduce these hazards. The aim of the present study is to evaluate the effects of the plant extract essential oil (EO) vapors of Neem, Clove, Cinnamon bark, Thyme, Lemon Grass, and Eucalyptus on the bacterial count in bioaerosols near dental units. Materials and Methods: Sampling was taken on nutrient blood agar plates by placing them open near dental units using passive air sampling method, before commencement of treatment for 1 h, during treatments for 2 h, and after introducing EO vapors for 2 h. The collected samples were taken for incubation at 37°C for 48 h. The colonies formed were counted in colony-forming units per cubic meter and taken for statistical analysis. Results: After comparing the obtained results, it was found that there was a significant reduction (P < 0.05) in the bacterial count for about 43% near the dental units after the introduction of the EO vapours. Conclusion: It is concluded that natural extracts like EOs can reduce bacterial contamination near dental units in the vapourized state, thereby reducing the health hazards in Dental Health Professionals.

Evaluation of effects of essential oil vapors on the bacterial count in bioaerosols

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Production and characterization of alpha-amylase from Aspergillus niger JGI 24 isolated in Bangalore

Evaluation of effects of essential oil vapors on the bacterial count in bioaerosols