scholarly journals Prospects of Biocatalyst Purification Enroute Fermentation Processes

2021 ◽  
Author(s):  
Michael Bamitale Osho ◽  
Sarafadeen Olateju Kareem
Keyword(s):  
Fermentation Process ◽  
Colloidal Particles ◽  
Methyl Cellulose ◽  
Industrial Applications ◽  
Maximum Activity ◽  
Inorganic Materials ◽  
Organic Substrates ◽  
Ammonium Sulphate Precipitation ◽  
Sodom Apple ◽  
Microbial Agents

Biotransformation of broth through fermentation process suffers a major setback when it comes to disintegration of organic substrates by microbial agents for industrial applications. These biocatalysts are in crude/dilute form hence needs to be purified to remove colloidal particles and enzymatic impurities thus enhancing maximum activity. Several contractual procedures of concentrating dilute enzymes and proteins had been reported. Such inorganic materials include ammonium sulphate precipitation; salting, synthetic polyacrylic acid; carboxy-methyl cellulose, tannic acid, edible gum and some organic solvents as precipitants etc. The emergence of organic absorbents such as sodom apple (Calostropis procera) extract, activated charcoal and imarsil had resulted in making significant impact in industrial circle. Various concentrations of these organic extracts have been used as purifying agents on different types of enzyme vis: lipase, amylase, protease, cellulase etc. Purification fold and stability of the enzyme crude form attained unprecedented results.

scholarly journals Review on Microbial Sources, Production, Purification and Potential Industrial Applications of Laccases

2021 ◽  
Vol 8 (12) ◽  
pp. 39-74
Author(s):  
Amit Rastogi ◽  
Rajesh Singh ◽  
Ahmed Barhoums
Keyword(s):  
Higher Plants ◽  
Inorganic Compounds ◽  
Industrial Applications ◽  
Exchange Chromatography ◽  
Organic Substrates ◽  
Electron Transfer Mechanism ◽  
Source Effect ◽  
Ammonium Sulphate Precipitation ◽  
Production Techniques ◽  
Pharmaceutical Industries

Laccase (EC 1.10.3.2) is a multicopper blue oxidase which are involved in the oxidation of a broad range of organic substrates, including phenols, polyphenols, anilines, and even certain inorganic compounds by a one-electron transfer mechanism. Laccases are widely distributed in bacteria, fungai, insects and higher plants. There are mainly two production techniques for cultivation of laccase such as submersed fermentation and solid- state fermentation. This paper briefly discuss the effect of carbon source, effect of nitrogen source, effect of inducers, effects of surfactants, effect of agitator, influence of metal ions and use of agro-industrial waste in production medium. The paper also discussed the purification techniques such as ammonium sulphate precipitation for extraction purpose followed by dialysis and ion-exchange chromatography as well characterization techniques. Laccases are known to show application ranging from pharmaceutical industries to textile sector as well as in biosensor development.

scholarly journals Man/Cel5B, a Bifunctional Enzyme Having the Highest Mannanase Activity in the Hyperthermic Environment

2021 ◽  
Vol 9 ◽  
Author(s):  
Beenish Sadaqat ◽  
Chong Sha ◽  
Parveen Fatemeh Rupani ◽  
Hongcheng Wang ◽  
Wanbing Zuo ◽  
...  
Keyword(s):  
Thermotoga Maritima ◽  
Methyl Cellulose ◽  
Industrial Applications ◽  
Maximum Activity ◽  
Gene Encoding ◽  
Genes Encoding ◽  
Mannanase Activity ◽  
Encoding Protein ◽  
Layer Chromatography ◽  
Carboxy Methyl

Thermotoga maritima (Tma) contains genes encoding various hyperthermophilic enzymes with great potential for industrial applications. The gene TM1752 in Tma genome has been annotated as cellulase gene encoding protein Cel5B. In this work, the gene TM1752 was cloned and expressed in Escherichia coli, and the recombinant enzyme was purified and characterized. Interestingly, the purified enzyme exhibited specific activities of 416 and 215 U/mg on substrates galactomannan and carboxy methyl cellulose, which is the highest among thermophilic mannanases. However, the putative enzyme did not show sequence homology with any of the previously reported mannanases; therefore, the enzyme Cel5B was identified as bifunctional mannanase and cellulase and renamed as Man/Cel5B. Man/Cel5B exhibited maximum activity at 85°C and pH 5.5. This enzyme retained more than 50% activity after 5 h of incubation at 85°C, and retained up to 80% activity after incubated for 1 h at pH 5–8. The Km and Vmax of Man/Cel5B were observed to be 4.5 mg/mL galactomannan and 769 U/mg, respectively. Thin layer chromatography depicted that locust bean gum could be efficiently degraded to mannobiose, mannotriose, and mannooligosaccharides by Man/Cel5B. These characteristics suggest that Man/Cel5B has attractive applications for future food, feed, and biofuel industries.

scholarly journals Culture-Independent and Culture-Dependent Characterization of the Black Soldier Fly Gut Microbiome Reveals a Large Proportion of Culturable Bacteria with Potential for Industrial Applications

2021 ◽  
Vol 9 (8) ◽  
pp. 1642
Author(s):  
Dorothee Tegtmeier ◽  
Sabine Hurka ◽  
Sanja Mihajlovic ◽  
Maren Bodenschatz ◽  
Stephanie Schlimbach ◽  
...  
Keyword(s):  
Gut Microbiome ◽  
Amplicon Sequencing ◽  
Industrial Applications ◽  
Culture Collection ◽  
Rrna Gene ◽  
Organic Substrates ◽  
Culturable Bacteria ◽  
Black Soldier Fly ◽  
Culture Independent ◽  
Culture Dependent

Black soldier fly larvae (BSFL) are fast-growing, resilient insects that can break down a variety of organic substrates and convert them into valuable proteins and lipids for applications in the feed industry. Decomposition is mediated by an abundant and versatile gut microbiome, which has been studied for more than a decade. However, little is known about the phylogeny, properties and functions of bacterial isolates from the BSFL gut. We therefore characterized the BSFL gut microbiome in detail, evaluating bacterial diversity by culture-dependent methods and amplicon sequencing of the 16S rRNA gene. Redundant strains were identified by genomic fingerprinting and 105 non-redundant isolates were then tested for their ability to inhibit pathogens. We cultivated representatives of 26 genera, covering 47% of the families and 33% of the genera detected by amplicon sequencing. Among these isolates, we found several representatives of the most abundant genera: Morganella, Enterococcus, Proteus and Providencia. We also isolated diverse members of the less-abundant phylum Actinobacteria, and a novel genus of the order Clostridiales. We found that 15 of the isolates inhibited at least one of the tested pathogens, suggesting a role in helping to prevent colonization by pathogens in the gut. The resulting culture collection of unique BSFL gut bacteria provides a promising resource for multiple industrial applications.

A novel Swollenin from Talaromyces leycettanus JCM12802 exhibits cellulase activity and enhanced cellulase hydrolysis towards various substrates

2020 ◽  
Author(s):  
Honghai Zhang ◽  
Yuan Wang ◽  
Roman Brunecky ◽  
Bin Yao ◽  
Xiangming Xie ◽  
...  
Keyword(s):  
Synergistic Effect ◽  
Functional Diversity ◽  
Specific Activity ◽  
Biomass Conversion ◽  
Hydrolytic Enzymes ◽  
Cellulase Activity ◽  
Industrial Applications ◽  
Maximum Activity ◽  
Carbohydrate Binding ◽  
Cell Wall Polysaccharides

Abstract Background Swollenins are present in some fungal species involved in the biodegradation of cellulosic substrates. They appear to promote a rearrangement in the network of non-covalent interactions between the cell wall polysaccharides, thus making it more accessible for degradation by hydrolytic enzymes. Here, we have reported a detailed characterization of a recombinant swollenin with respect to its disruptive activity on cellulosic substrates and synergistic effect with cellulases. Results In the present study, a novel swollenin gene Tlswo consisting of an open reading frame encoding 503 amino acids was identified from Talaromyces leycettanus JCM12802 and successfully expressed in Trichoderma reesei and Pichia pastoris. Similar to other fungal swollenins, TlSWO contained a N-terminal family 1 carbohydrate binding module (CBM1) followed by a Ser/Thr rich linker connected to expansin-like domain which includes a family 45 endoglucanase-like domain and group-2 grass pollen allergen domain. TlSWO demonstrated disruptive activity on Avicel and displayed a high synergistic effect with cellobiohydrolases, enhancing its hydrolytic performance up to 132%. The activity of TlSWO on various substrates and biomass was also examined. It was shown that TlSWO could release reducing sugars from lichenan, barley β-glucan, carboxymethyl cellulose sodium (CMC-Na) and laminarin. The specific activity of TlSWO towards the substates above is 9.0 ± 0.100 U/mg, 8.9 ± 0.100U/mg, 2.3 ± 0.002 U/mg and 0.79 ± 0.002 U/mg respectively. Moreover, TlSWO exhibits maximum activity at pH 4.0 and 50 ℃. Conclusion This study reported on a novel swollenin with highly efficient for biomass conversion. It also reveals the functional diversity of swollenin with activity on various substrates. Although the exact mechanism of swollenin catalytic action activity still remains unknown, the functional diversity of TlSWO makes it a good candidate for industrial applications.

scholarly journals Laundry Detergent Compatibility of Papain Like Protease Purified From Piper Betel Leaves

2018 ◽  
Vol 7 (3.3) ◽  
pp. 132
Author(s):  
A Mousami Shankar ◽  
Dr G.V.D. Sirisha ◽  
Dr K. Vijaya Rachel
Keyword(s):  
Laundry Detergent ◽  
Deae Cellulose ◽  
Maximum Activity ◽  
Native Page ◽  
Chromatographic Techniques ◽  
Detergent Compatibility ◽  
Ammonium Sulphate Precipitation ◽  
Sds Page ◽  
The Stability ◽  
Better Than

Enzymes have wide applications in detergent industry from early 1900’s. Mostly, clothes are soiled by protein based grime. Most of the detergents have either amylase / protease. Various sources were scrutinized for potent protease activity and Betel leaves were selected, the enzyme was then isolated, purified to homogeneity by ammonium sulphate precipitation, DEAE-Cellulose and gel permeation chromatographic techniques. The enzyme was monomeric in nature with a molecular mass of 38kDa as determined by native PAGE and SDS-PAGE. The enzyme shows maximum activity at 60oC and pH 4.0. The Km and Vmax of the enzyme were 4x10-3M and 54µmol/min/mg respectively. The enzyme was categorically inhibited by PCMB and iodo-acetamide suggesting it to have papain like nature. The stability of the enzyme is assessed over the stretch of alkaline pH and temperature. This evaluation validates the stability of the enzyme and its use in detergent formulations. It was evident that after adding the enzyme preparation the stains (tea, chocolate, blood) were removed much better than that of the controls, which affirms that papain like enzyme from betel leaves, enhances detergent activity.  

scholarly journals Characterization of a Novel β-Xylosidase, XylC, fromThermoanaerobacterium saccharolyticumJW/SL-YS485

2010 ◽  
Vol 77 (3) ◽  
pp. 719-726 ◽  
Author(s):  
Weilan Shao ◽  
Yemin Xue ◽  
Ailian Wu ◽  
Irina Kataeva ◽  
Jianjun Pei ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Glycosyl Hydrolase ◽  
Gel Filtration ◽  
Industrial Applications ◽  
Maximum Activity ◽  
Site Directed Mutagenesis ◽  
Stem Loop ◽  
Translational Initiation ◽  
Reading Frame ◽  
Gradient Gel Electrophoresis

ABSTRACTThe 1,914-bp open reading frame ofxylCfromThermoanaerobacterium saccharolyticumJW/SL-YS485 encodes a calculated 73-kDa β-xylosidase, XylC, different from any glycosyl hydrolase in the database and representing a novel glycohydrolase family. Hydrolysis occurred under retention of the anomeric configuration, and transglycosylation occurred in the presence of alcohols as acceptors. With the use of vector pHsh, expression of XylC, the third β-xylosidase in this bacterium, increased approximately 4-fold when a loop within the translational initiation region in the mRNA was removed by site-directed mutagenesis. The increased expression ofxylCmis due to removal of a stem-loop structure without a change of the amino acid sequence of the heterologously expressed enzyme (XylCrec). When gel filtration was applied, purified XylC had molecular masses of 210 kDa and 265 kDa using native gradient gel electrophoresis. The protein consisted of 78-kDa subunits based on SDS gel electrophoresis and contained 6% carbohydrates. XylC and XylCrecexhibited maximum activity at 65°C and pH65°C6.0, a 1-h half-life at 67°C, aKmforp-nitrophenyl-β-d-xyloside of 28 mM, and aVmaxof 276 U/mg and retained 70% activity in the presence of 200 mM xylose, suggesting potential for industrial applications.

scholarly journals Improvement of Biogas Yield by Pre-Treating Poultry Waste with Bacterial Strains

Energies ◽  
2021 ◽  
Vol 14 (18) ◽  
pp. 5601
Author(s):  
Krystyna Cybulska ◽  
Ilona Kołosowska ◽  
Karol Kramkowski ◽  
Monika Karpińska ◽  
Katarzyna Roszkowicz-Ostrowska ◽  
...  
Keyword(s):  
Environmental Sustainability ◽  
Fermentation Process ◽  
Industrial Applications ◽  
Biogas Plant ◽  
Meat Industry ◽  
Actual Process ◽  
Poultry Waste ◽  
Bacterial Strains ◽  
Methane Fermentation ◽  
Biological Sludge

Poultry waste is increasingly used as a substrate for the methane fermentation process in a biogas plant. However, not all waste materials processed in the meat industry meet the criteria for optimal process management and cost-effective methane efficiency. An example may be centrifuged biological sludge, etc. Treatment of such material used as a substrate by introducing new metabolically and enzymatically active strains of bacteria could be beneficial for the fermentation process in a biogas plant and provide increased energy efficiency. The aim of the study was to compare the amount and quality of biogas obtained from biological sludge from the processing of poultry vaccinated with metabolically diversified bacterial inoculum after initial incubation of the batch before the actual process in a biofermenter. Laboratory tests were carried out in accordance with the guidelines contained in the DIN 38 414-S8 and VDI 4630 standards. Based on the obtained results, it was found that the optimized biological sludge can be used as a substrate in the methane fermentation process in a biogas plant. The material processed by the combination of bacterial strains marked with the symbols A/C, E/G, and F/H showed a significantly increased efficiency of biogas, including methane, compared to the non-grafted material. This is a good predictor for industrial applications, process feasibility, economic viability, and environmental sustainability that should be compiled based on the results obtained from this study.

Industrial applications of Raman microscopy

1992 ◽  
Vol 50 (2) ◽  
pp. 1502-1503
Author(s):  
D.L. Gerrard
Keyword(s):  
Laser Scanning ◽  
Near Infrared ◽  
Industrial Applications ◽  
Raman Microscopy ◽  
Inorganic Materials ◽  
X Ray ◽  
Spatially Resolved ◽  
Wide Range ◽  
Recent Developments ◽  
Scanning Optical Microscopy

One of the major advantages of Raman spectroscopy for the industrial analyst is its capability for providing spatially resolved molecular information on a wide range of inorganic materials. Although the technique of Raman microscopy has been available for nearly twenty years its value in industrial analysis is still not widely appreciated. Recent developments in the use of near infrared excitation with Fourier transform spectrometers and of microline focus systems with charge-coupled devices as detectors have greatly expanded the value of the technique and should help it to appeal to a wider audience. Raman microscopy provides much valuable information in its own right and can often be used to solve analytical problems without reference to any other technique. However, it is usually of greatest value to the industrial analyst when used in conjunction with other microspectroscopy techniques such as scanning electron microscopy/energy dispersive X-ray analysis, infrared microscopy, proton-induced X-ray emission, laser ionisation mass analysis and laser scanning optical microscopy.

Novel Wet Chemical Copper Metallization for Glass Interposers

2011 ◽  
Vol 2011 (DPC) ◽  
pp. 001535-001554
Author(s):  
Simon Bamberg ◽  
Ralf Bruening ◽  
Johannes Etzkorn ◽  
Frank Bruening
Keyword(s):  
Manufacturing Industry ◽  
Copper Layer ◽  
Inorganic Materials ◽  
Copper Metallization ◽  
Organic Substrates ◽  
Copper Plating ◽  
Electrolytic Copper ◽  
Electroless Copper ◽  
Wet Chemical ◽  
The Impact

The focus of the chip and IC substrate manufacturing industry for interposers is currently shifting from organic substrates to inorganic materials. Interposers overcome the dimensional mismatch between a die and an organic PWB substrate and need to buffer the differences in thermal expansion between these two materials. While inorganics like silicon and glass, have a low CTE value compared to organic material, glass has some significant advantages over silicon. These are namely material cost, availability in panel size and a better electrical insulation. In order to further increase the cost advantage, metallization could be done by either electroless or electrolytic copper plating as an alternative route to sputter technology. Both wet chemical processes are well established in PWB manufacturing and need adaptation to glass substrates. Compared to sputtering, 3D-features can be covered with a wet chemical treatment in a comparatively economical process. Filling of TGVs (Through Glass Vias) by electrolytic copper plating requires the presence of a conductive film which is created by applying electroless copper deposition of typically 300–500nm thickness. The general issue is poor adhesion to the smooth glass surfaces. To improve adhesion between glass and metal, various treatments were assessed in this study: Mechanical anchoring was achieved by etching the substrate (subtractive) as well as by depositing nano-sized silica particles (additive). Both treatments are in solution and 3D-features are accessible in contrast to a purely mechanical approach. Surface treatments without inducing additional roughness included the adsorption of functional metal-affine polyelectrolytes and silanization for conditioning which enhanced the adhesion of the copper layer to various degrees. To study the impact of chemical formulation on strain/stress development and relaxation in the film as a key impact factor for blister (adhesion) performance, the electroless copper layer growth was monitored by in-situ XRD methods during and after deposition.

Characterization of L-fucose isomerase from Paenibacillus rhizosphaerae to produce L-fuculose from hydrolyzed fucoidan and commercial fucose

2019 ◽  
Author(s):  
Muhammad Waheed Iqbal ◽  
Tahreem Riaz ◽  
Shahid Mahmood ◽  
Yingying Zhu ◽  
Dawei Ni ◽  
...  
Keyword(s):  
Industrial Applications ◽  
Maximum Activity ◽  
Lysosomal Disease ◽  
Simple Method ◽  
Enzymatic Production ◽  
Native Molecular Mass ◽  
Anti Cancer ◽  
Viral Hepatitis B ◽  
High Production

Abstract Background L-fuculose is an expensive and rare sugar used against different kinds of diseases such as HIV, anti-cancer, anti-viral, Hepatitis-B, human lysosomal disease (fucosidosis), and cardio-protective drugs. The enzymatic way of converting L-fucose into L-fuculose would be an effective method with great industrial applications. The purpose of this research is to introduce a high production of L-fuculose from cheap and natural sources (fucoidan) and commercially source (Sigma-Aldrich) by a recombinant enzyme L-fucose isomerase from Paenibacillus rhizosphaerae (Pa-LFI).Results Fucose containing polysaccharide (FPs) called fucoidan was extracted, hydrolyzed and characterized by U. pinnatifida for enzymatic production of L-fuculose. The FPs provide 35.9% of fucose along with few other monosaccharides. Pa-LFI was characterized and purified with a single band at 65 kDa. It showed an activity of 104.5 U mg -1 and exhibited as a hexamer with native molecular mass 396 kDa. The maximum activity for recombinant Pa-LFI was detected at pH 6.5 and 50 °C in 1 mM of Mn 2+ . The melting temperature observed 75 °C and half-life at 50 °C was 12.6 h. The isomerizing activity of Pa-LFI with aldose substrate (L-fucose) was higher exposing K m , k cat and k cat / K m 86.2 mM, 32831 min -1 and 335 min -1 mM -1 respectively. The conversion ratio of L-fuculose from 100 g L -1 of FPs and commercial fucose after the equilibrium state was about 6% (5.6 g L -1 ) and 30% (30.2 g L -1 ) respectively.Conclusion Pa-LFI catalyzed the reaction to convert L-fucose into L-fuculose. The enzyme will be helpful in the production of L-fuculose with an efficient and simple method without producing any by-product.

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