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The consumption of fuel for transportation is increasing during the last decade. Bioethanol is one of the renewable energy has a good opportunity to be applied when the lack of fossil fuel. Bioethanol is derived from the lignocellulose substance through a fermentation process. In this research, the lignocellulose came from the petai peel (Parkia speciosa). The peel was hydrolyzed using an enzyme and continuously fermented for 5 days. The aim of this research is to analyst the mass balance of the bioethanol production from petai peel (Parkia speciosa) through the enzymatic process. The enzyme used in this research are alfa amylase (10 ml) and glucoamylase (10 ml), also Saccharomyces cerevisiae used in the fermentation process. The result shows that the initial material of petai peel was 57 grams will produce bioethanol around 14 grams.

Objectives Platelet-rich fibrin (PRF) has been utilized in regenerative medicine as a concentration of autologous platelets and growth factors that stimulates tissue regeneration. More recently, liquid-PRF (also called injectable-PRF; i-PRF) has been brought to market utilizing PET plastic tubes. Due to new advances made in tube technology, the first aim of the present study was to investigate the liquid consistency of liquid-PRF utilizing both standard and chemically modified PET plastic tubes. Furthermore, it is well known that the conversion of PRF into a fibrin matrix is derived from the temperature-controlled enzymatic process that converts liquid fibrinogen and thrombin to solid fibrin. This study also investigated for the first time the use of a cooling device (Bio-Cool) to extend the liquid working properties of liquid-PRF. Materials and methods In total, 30 participants enrolled in this study. From each patient, four tubes of liquid-PRF were drawn, two standard white Vacuette tubes and two blue chemically modified hydrophobic tubes. Following centrifugation at 700 RCF-max for 8 min in a Bio-PRF horizontal centrifuge, one white and one blue tube were kept upright at room temperature, while the other white and blue tube were placed within the cooling device. Thereafter, the liquid-PRF layers were monitored over time until clotting occurred. Patient gender, age, and altitude above sea level (+ 5000 ft) were recorded and compared for clotting times. Results The findings from the present study demonstrated that the chemically modified PET tubes performed 37% better than the control tubes (extended the working properties of liquid-PRF by over 20 min). Most surprisingly, tubes kept in the cooling device demonstrated an average of 90 min greater working time (270% improvement). While patients living at altitude did significantly improve the clotting ability of liquid-PRF, no differences were observed when comparing male vs female or younger vs older patients in liquid-PRF clotting times. Conclusions Cooling of blood following centrifugation represented a 270% improvement in working properties of liquid-PRF. Optimization of liquid-PRF tubes utilizing chemically modified hydrophobic PET tubes also delayed the clotting process by 37%. Patient gender and age had little relevance on liquid-PRF. Clinical relevance The present findings demonstrate for the first time that cooling of liquid-PRF is able to extend the working properties of liquid-PRF by over 90 min. Thus for clinicians performing longer clinical procedures, the cooling of blood may represent a viable strategy to improve the working time of liquid-PRF in clinical practice.

Variable industrial strains have been applied in the fermentation of bulk amino acids. Strain discovery and evolution, process optimization are traditional approaches to improve the yield and efficiency of the bio-production process, hence to compete with chemical or enzymatic process in amino acids production. With the fast development of bioengineering and synthetic biology, the strains can be rationally engineered to achieve better performance and gain the capacity in the fermentation of broader range of amino acids, especially for value-added amino acids. This proposed review aims to summarize traditional and recent strains in the microbial production of amino acids, characterize their metabolic pathways and present potential objectives for rational evolution. In addition, this proposed review prospect the recent opportunities and challenges in the microbial production of value-added amino acids (rare amino acids, non-canonical amino acids, and unnatural amino acids).

Background Designer lipid is a novel, health-friendly lipid with potential application in food, nutraceutical and pharmaceutical industries, including obesity, cancer, heart disease, inflammation. These advantages arise due to modification of fat/oil by the chemical or enzymatic process to form designer lipid. The transformation of lipid results in the rearrangement of fatty acid within a triglyceride molecule or between two different triglycerides. Thus, the resulting designer lipid has superior and unique physicochemical properties than the naturally occurring triglycerides. Due to these excellent physicochemical properties, they are in great demand in the market. Scope and approach The primary aim of this review is to describe component fatty acids used for the synthesis of designer lipids, the process used in designing designer lipids, and reactors used to intensify the yield of designer lipids, application of designer lipid in the food and nutraceutical sector. Key findings and conclusions Designer lipid is a chemically/enzymatically modified form of lipid to improve physicochemical and nutritional properties of traditional lipids coming from plant and animal source. Such fabricated lipids have attracted consumers' attention because of their unique properties and capability to manage various syndromes. Their demand by the consumer increased over the recent past. To fulfil the increase in demand, the intensified synthesis of these designer lipids is carried out using packed bed reactors, ultrasonic reactors, high-pressure reactors. With their claimed health benefits, designer lipids are widely used as a functional ingredient in the food and pharmaceutical industries. Currently, designer lipids are used as a plastic fat, human milk substitute, cocoa butter, used in infant formulation, low-calorie lipids, an anti-cancer, reduced cardiovascular risk etc. The application of designer lipid is governed by the positional distribution and type of fatty acid esterified on the glycerol backbone.

Multi-enzymatic reactions can successfully replace complex chemical syntheses, using milder reaction conditions, and generating less waste. The present model-based analysis compares the performances of several optimally operated Batch Reactors (BR) with those of an optimally operated serial Sequence of BRs (SeqBR). In multi-enzymatic systems, SeqBR could be more advantageous and flexible, allowing the optimization of costly enzymes amounts used in each BR in the series. Exemplification was made for the bi-enzymatic reduction of D-fructose to mannitol by using MDH (mannitol dehydrogenase) and the NADH cofactor, with the in situ continuous regeneration of NADH at the expense of formate degradation in the presence of FDH (formate dehydrogenase). For such coupled enzymatic systems, the model-based engineering evaluations are difficult tasks, because they must account for the common species’ initial levels, their interaction, and their dynamics. The determination of optimal operating modes of sole BR or of a SeqBR turns into a multi-objective optimization problem with multiple constraints to be solved for every particular system. The study presents multiple elements of novelty: (i) the proof of higher performances of an optimal SeqBR (including N-BRs) compared to a sole optimal BR operated for N-number of runs and (ii) the effect of using a multi-objective optimization criteria on SeqBR adjustable dynamics.

Exopolysaccharides (EPSs) are classified in two groups, homopolysaccharides (HoPs) and heteropolysaccharides (HePs), are produced by lactic acid bacteria (LAB) and used in a range of industrial applications including the medical and food industries. HoPs are extracellular EPSs and their production depends on extracellular enzymes, while HePs are intracellular EPSs. Their nature (extra or intracellular) directly impacts production rates, HoPs having higher yields. The development of processes for producing EPSs has attracted great interest, since novel application trends have emerged due to the great diversity of recent information generated on EPSs properties. HoPs have been synthesized by fermentation using bacterial cells and a cell-free enzymatic process, while HePs have been produced only by fermentation. The analysis of the EPSs production processes indicates that macronutrients such as the carbon and nitrogen source used in the culture media are very important for the synthesis of EPSs and the enzymes involved, understanding their importance can assist to design processes for production of EPSs with desirable characteristics and yields according to the needs of the processes and products to which they are applicable. This review emphasized in the analyses of carbon and nitrogen sources used for EPSs production and their functional applications and productive aspects.

Currently, the growth of diseases with diabetes, metabolic disorders, and obesity increases the demand for preventive and functional products. Currently available technologies for the production of preventive products provide for the replacement of sugar with sugar substitutes or its complete absence, which naturally reduces the nutritional and energy value of the products produced. One of the main promising areas of the processing industry is the production of new products and sugar substitutes based on non-traditional types of plant raw materials that have a rich carbohydrate complex in their composition. These types of raw materials among vegetable crops include jerusalem artichoke. Currently, jerusalem artichoke juice is of particular interest for use in the production of soft drinks, as it has a rich chemical composition, which makes Jerusalem artichoke indispensable in dietary nutrition, in the preparation of highly effective medicines. In this article, the chemical composition of tubers and juice-semi-finished products from jerusalem artichoke is investigated. The results of the studies showed that the bulk of the dry substances in jerusalem artichoke tubers are carbohydrates, most of which are represented by fructosides. In the semi-finished juice passes: oligosaccharides 22.54 %, monosaccharides 1.61%, as well as part of the structural polysaccharides-fiber 0.07 %, pectin substances 1.84 %. The change in the fractional composition of the polyphenolic substances of the pulp and semi - finished juice was studied when the pulp was kept for 30 minutes. When developing the technology for the production of juices and beverages based on jerusalem artichoke, it was found that the polyphenolic substances of jerusalem artichoke tubers have a great influence on the technological properties of raw materials, the quality and nutritional value of the finished product. It was found that the enzymatic process of oxidative transformation of polyphenols occurs as much as possible in the first 5-10 minutes.

Enzymatic biodiesel production has been at the forefront of biofuels research in recent decades because of the significant environmental advantages it offers, while having the potential to be as effective as conventional chemically catalyzed biodiesel production. However, the higher capital cost, longer reaction time, and sensitivity of enzyme processes have restricted their widespread industrial adoption so far. It is also posited that the lack of research to bring the biodiesel product into final specification has scuppered industrial confidence in the viability of the enzymatic process. Furthermore, the vast majority of literature has focused on the development of immobilized enzyme processes, which seem too costly (and risky) to be used industrially. There has been little focus on liquid lipase formulations such as the Eversa Transform 2.0, which is in fact already used commercially for triglyceride transesterification. It is the objective of this review to highlight new research that focuses on bringing enzymatically produced biodiesel into specification via a liquid lipase polishing process, and the process considerations that come with it.

Efficient saccharification of lignocellulosic biomass requires concerted development of a pretreatment method, an enzyme cocktail and an enzymatic process, all of which are adapted to the feedstock. Recent years have shown great progress in most aspects of the overall process. In particular, increased insights into the contributions of a wide variety of cellulolytic and hemicellulolytic enzymes have improved the enzymatic processing step and brought down costs. Here, we review major pretreatment technologies and different enzyme process setups and present an in-depth discussion of the various enzyme types that are currently in use. We pay ample attention to the role of the recently discovered lytic polysaccharide monooxygenases (LPMOs), which have led to renewed interest in the role of redox enzyme systems in lignocellulose processing. Better understanding of the interplay between the various enzyme types, as they may occur in a commercial enzyme cocktail, is likely key to further process improvements.

DNA double-strand break (DSB) resection, once thought to be a simple enzymatic process, is emerging as a highly complex series of coordinated activities required to maintain genome integrity. Progress in cell biology, biochemistry, and genetics has deciphered the precise resecting activities, the regulatory components, and their ability to properly channel the resected DNA to the appropriate DNA repair pathway. Herein, we review the mechanisms of regulation of DNA resection, with an emphasis on negative regulators that prevent single-strand (ss)DNA accumulation to maintain genome stability. Interest in targeting DNA resection inhibitors is emerging because their inactivation leads to poly(ADP-ribose) polymerase inhibitor (PARPi) resistance. We also present detailed regulation of DNA resection machineries, their analysis by functional assays, and their impact on disease and PARPi resistance.

The past decade has witnessed a spectacular growth in bioproducts development due to the intense interest in creating a more diverse energy supply and a carbon neutral bioeconomy. Such a burgeoning...

The enzymatic carbon fixation is a promising approach to deal with greenhouse gas emission and is usually accompanied with energy consumption during the reduction of CO2. As a very important route, the carboxylation can convert CO2 to organic carbon without extra requirement of reduction power and is hoped as a greener solution, especially for some non-bulk chemicals, such as medical intermediates. Here, a concept-proof trail of green enzymatic process of conversing both of CO2 and benzene to produce 2,3-dihydroxybenzoic acid (2,3-DHBA), which is the intermediate for fine chemicals, is introduced with O2 from air and glucose. The results showed that the conversion catechol by 2, 3-dihydroxybenzoic acid decarboxylase (2,3-DHBD) alone was around 30 %, with an overall conversion from phenol of 2.4 %, which was limited by the in-situ production of catechol. This trail contributed a green enzymatic route for the production of 2,3-DHBA.

Lead is generated as residual progress of vehicle or factory process. The excessive amount of lead has several impacts on human health system namely on cardiovascular, respiratory, endocrine and metabolic system through damaging enzymatic process and Reactive Oxygen Species (ROS). The aim of this study is to search the correlation of lead exposure on blood pressure and lipid parameters alteration among people around bus terminal in Yogyakarta. This study is a cross sectional research design. According to calculation of sample size, 71 respondents had fulfilled the inclusion criteria such as: Agree to be respondents, 6 hours fasting prior to take data, age >18 years old and had history of staying nearby bus terminal for more than one year. The variables divided into two, independent variable is lead exposure and dependent variables are SBP and DBP meanwhile lipid parameters such as LDL, HDL and TG. Those variables were analyzed by linier regression. Based on linier regression was obtained that SBP (p value 0.004, r 0.33 and R square 0.11), DBP (p value 0.001 r 0.37 and R square 0.138), LDL (p value 0.002 r 0.35 and R square 0.128), HDL (p value 0.001, r 0.37 and R square 0.139) and TG (p value 0,002, r 0.36 and R square 0.130). The result of study was found that lead exposure contributed on blood pressure and lipid parameters alteration among people around bus terminal in Yogyakarta.

In biofuel/biochemical production, starch is the current most used feedstocks while lignocellulosic biomass is a more economical, non-food competitive alternate. This chapter discussed in details on enzymes that are used to break down these to feedstocks: amylases for starch breaking down; cellulases, hemicellulases, and lignin-related enzymes for lignocellulosic biomass degrading. Their functions, structures, mechanisms, and productions were all discussed. The applications of commercially available enzymes for the conversion of biomass feedstocks into biofuels and bioproducts in food, feed, biopharmaceutical industries were also addressed. To reduce the shortcomings of the enzymatic process such as high cost, the cascade biorefinery process for a complete valorization of biomass or processing wastes could enhance the economy and sustainability of the process.

Enzymatic interventions in animal skin processing are increasingly being considered as safe and benign technology options due to the reduction and replacement of potential harmful chemicals. In this study, galactosidases have been employed for rehydration of preserved skins and hides to improve the process efficiency and minimize hazardous sodium sulfide. The purpose of rehydration is to ensure the skin is hydrated uniformly to facilitate subsequent physico-chemical processes of leather making. Improper rehydration leads to reduction in the quality and value of the leather. The efficacy of the enzymatic process was studied using histological images and scanning electron microscopic analysis. Pollution load changes and the extent of carbohydrate removal were also quantified. The study indicates possibility for substantial reduction in process duration and water input (up to 30%) during rehydration of preserved animal skins when galactosidases are used as rehydration aid without affecting the quality of the leather. Thus use of galactosidases in rehydration ensures uniform accelerated rehydration and provides significant environmental benefits to tanning industry, by reducing harmful substances in subsequent operations.

In this study, deep eutectic solvent (DES) used as co solvent for enzymatic biodiesel production from degumming palm oil (DPO). DES is formed from the salt compound choline-chloride (ChCI) with glycerol at 1:2 molar ratio. Furthermore, the effectiveness of the DES was tested by enzymatic reactions using novozym 435® for the production of palm biodiesel with raw materials DPO. The use of enzymes with the DES system can maintain the activity and stability of the novozyme enzyme measured by the yield produced until the 10th usage produces biodiesel yield > 99% with a concentration of DES 0.5% with a molar ratio of 0.5% water. Spectra of DES ChCI:glycerol and ChCI:glycerol:water characterized by FTIR, morphological structure novozym by characterized SEM and then biodiesel product analyzed by GC-MS. This shows that the ChCI:glycerol:water system in enzymatic biodiesel production has good potential to maintain enzyme activity and stability.

We introduce a recyclable and efficient enzymatic process by integration of microwave (MW) pretreatment followed by magnetic nanobiocatalysis for the green and sustainable oilseed biorefinery. The ...

The specificity of fructooligosaccharides as prebiotics depends on their size and structure, which in turn depend on their origin or the synthesis procedure. In this work we describe the application of an inulosucrase (IslA) from Leuconostoc citreum CW28 to produce high molecular weight inulin from sucrose alongside a commercial endoinulinase (Novozym 960) produced by Aspergillus niger for a simultaneous or sequential reaction to synthesize fructooligosaccharides (FOS). The simultaneous reaction resulted in a higher substrate conversion and a wide diversity of FOS when compared to the sequential reaction. A shotgun MS analysis of the commercial endoinulinase preparation surprisingly revealed an additional enzymatic activity: a fructosyltransferase, responsible for the synthesis of FOS from sucrose. Consequentially, the range of FOS obtained in reactions combining inulosucrase from Ln. citreum with the fructosyltransferase and endoinulinase from A. niger with sucrose as substrate may be extended and regulated.

(S)-2-Chloro-1-(2,4-dichlorophenyl) ethanol (3) is a chiral intermediate in the synthesis of luliconazole ((R)-E-1). Here, we report a novel biopreparation of 3 by bioreduction of 2-chloro-1-(2,4-d...

The aim of this study was to analyze the effects of a rotating magnetic field (RMF) on the activity and stability of horseradish peroxidase (HRP) and the catalyzed enzymatic process. It was found that 8 h of exposure to RMF does not influence the operational stability of HRP. At the same time, it was observed that the RMF improved HRP activity in a wider range of pH: 49% and 68% at pH 6.5 and pH 7.0, respectively. The efficiency of the catalyzed enzymatic process under exposure to RMF was analyzed by two-dimensional correlation analysis (2DCorr) on the example of the Remazol Brilliant Blue R (RBBR) decolorization process. The obtained results indicated the influence of RMF on the intensification of this process. Our studies showed an alternative method of enzymatic process intensification, which is significant in science and has practical value for applications.

Background: Mathematical modeling is useful in the analysis, prediction, and optimization of an enzymatic process. Unlike the conventional modeling methods, Monte Carlo method has special advantages in providing representations of the molecule’s spatial distribution. However, thus far, Monte Carlo modeling of enzymatic system is namely based on unimolecular basis, not suitable for practical applications. In this research, Monte Carlo modeling is performed for enzymatic hydrolysis of lactose for the purpose of real-time applications. Results: The enzyme hydrolysis of lactose, which is conformed to Michaelis–Menten kinetics, is modeled using the Monte Carlo modeling method, and the simulation results prove that the model predicts the reaction kinetics very well. Conclusions: Monte Carlo modeling method can be used to model enzymatic reactions in a simple way for real-time applications.How to cite: Gao L, Guo Q, Lin H, et al. Modeling of lactose enzymatic hydrolysis using Monte Carlo method. Electron J Biotechnol 2019;41.https://doi.org/10.1016/j.ejbt.2019.04.010 Keywords: β-Galactosidase, Enzymatic hydrolysis, Enzymes, Galactose, Hydrolysis, Immobilized, Lactose, Modeling, Monte Carlo, Silica gel, Stochastic, Tagatose

BACKGROUND: The enzymatic process in continuous flow reactors has become attractive owing to the merits of in‐line purification, improved mixing and mass transfer, etc. However, the construction of continuous flow reactors requires stable and surface‐active monolithic scaffolds for enzyme immobilization. RESULTS: In this study, nanofibers of linear polyethyleneimine (LPEI) are formed on the surface of poly(sodium 4‐styrenesulfonate) (PSS)‐treated cordierite honeycomb (named LPEI/PSS cordierite‐H) as monolithic scaffold for enzyme immobilization. The controlled self‐assembly of LPEI ensures the formation of nanofibers (diameters of 20–80 nm) on the surface of cordierite‐H, which roughens the surface and offers many active sites for enzyme loading. The electrostatic interaction between negatively charged enzyme and positively charged LPEI helps the enzyme firmly attach on the surface. The loading capacity of LPEI/PSS cordierite‐H was 1.24 mg g⁻¹. Horseradish peroxidase (HRP)‐loaded LPEI/PSS cordierite‐H (HRP@LPEI/PSS cordierite‐H) exhibited activity recovery of 55.33%, 2‐fold higher than that of HRP‐loaded branched polyethyleneimine (BPEI)/PSS cordierite‐H (HRP@BPEI/PSS cordierite‐H) (23.72%). Furthermore, a silica layer was formed on the surface of HRP@LPEI/PSS cordierite‐H through LPEI‐induced silicification, which could prevent the enzyme from deactivation under extreme conditions. CONCLUSION: These monolithic biocatalysts have great potential for applications in wastewater treatment and even in other fields such as pharmaceutical synthesis. © 2019 Society of Chemical Industry

Aim Congenital disorders of glycosylation (CDG) belong to an expanding group of rare genetic metabolic disorders caused by defects in the complex chemical enzymatic process of glycosylation. The study is aimed at presenting a case report of a premature dysmorphic newborn, clinical presentation of the condition, the way it was diagnosed and treated, as well as its comparison with the known cases. Results The result of glycan analysis supports the assumption of a supposed glycosylation disorder and also specifies a specific subtype: CDG-1, subtype ALG12-CDG (Ig). Conclusion CDG have an extremely wide clinical spectrum and should be considered in any child with unexplained developmental delay, failure to thrive, seizures, and abnormalities in liver enzymes, coagulation and immunologic factors. The treatment of most forms of CDG depends upon numerous factors such as specific symptoms present, severity of the disorder, age and overall health of the patients and tolerance to certain medications or procedures. For these reasons, the treatment is specific for every individual. It is based on the symptoms and requires a coordination of efforts of a team of specialists (Tab. 4, Fig. 3, Ref. 19).

Enzymology is an important part of biotechnology. Its application range covers various industries such as food, pharmaceutical, agricultural, and chemical industries. Therefore, interest in the research field of enzyme has grown continuously and rapidly. A large number of processes in sugar industry have been reported to be catalyzed by commercially available enzymes. Although enormous progress in enzyme industrialization, the widespread application of enzymes for the functional carbohydrates production are still not fully achieved. To speed up the development of enzymatic production of functional carbohydrates, further research on several topics should be done to reduce the final operating costs for the industrial enzymatic process. In this chapter, the status and trends of enzyme engineering and immobilization of enzyme will be extensively discussed. These two technologies whether to modify the catalytic characteristic or to improve the reusability of industrial enzymes are hotspot issues to break through the bottlenecks in enzymology.

A new original application for a polyacrylic film based on the monomers 2-(dimethylamino)ethyl methacrylate (NNDA), 2-hydroxyethyl acrylate (2HEA) and methylmethacrylate (MMA) as a starch azure container has been set up for a simple determination of honey diastase activity. The proposed method is based on the correlation of reducing sugars generated during the enzymatic process with the Schade reference assay. The polyacrylic film is charged with starch azure acting as a container for this substance; thus, the starch does not interfere in the measurement of reducing sugars, so that the diastase activity is easily calculated. The method has been contrasted with Schade method, showing good correlation and differences under 0.4% between methods in some honey samples. The polyacrylic film has great potential for the routine honey diastase activity assessment in small laboratories, dramatically reducing analysis time and cost., FEDER (Fondo Europeo de Desarrollo Regional), and both the Spanish Ministerio de Economía, Industria y Competitividad (MAT2017-84501-R) and the Consejería de Educación—Junta de Castilla y León (BU306P18 and BU041G18)

Polyamines and polyamides have a fundamental role in the biology of plants, and the presence of NO seems compulsory to account for their actions. In general, the NO production has claimed to occur through an enzymatic process, but not involving polyamines and polyamides. Nevertheless, a non-enzymatic mechanism, such as an electron transfer process among polyamines or polyamides and an acid nitrite solution, could account for rapid production of NO, even in anoxic conditions. EPR experiments, carried out with these substrates, proved the formation of NO. This evidence supports a non-enzymatic mechanism as an alternative source of NO, even in plants. So, since the NO production seems directly dependent on polyamines or polyamides presence, and these responsible for many activities in plants, it comes plausibly to consider crucial the involvement of NO in their actions. Furthermore, as for mammals, these results would confirm that, even in plants, NO production can occur through both enzymatic and non-enzymatic mechanisms.

The small ubiquitin-like modifier (SUMO) is an important post-translational modifier that regulates various cellular processes. Extensive investigations have been made to comprehend the enzymatic process and consequence of SUMOylation. In vitro SUMOylation assays are invaluable for understanding the fundamental mechanisms of SUMOylation. A majority of these assays monitor changes in the size of the substrate upon SUMO conjugation. Current methods typically detect the size difference through SDS-PAGE and western blots, which makes these methods cumbersome, error-prone, and time-consuming. Here, we describe a fluorescence-based assay for real-time detection of SUMOylation. In the method, a fluorophore-tagged substrate is used in the SUMOylation reaction. Upon SUMOylation, the size and correlation time (τc ) of the substrate increases, and so does its anisotropy. The rate of change in anisotropy with time reflects the efficiency of the SUMOylation machinery. The real-time SUMOylation assay protocol is elegant, time-saving, and less prone to errors. © 2020 Wiley Periodicals LLC. Basic Protocol: Fluorescent anisotropy-based in vitro SUMOylation assay.

The removal of direct yellow 106 (DY106, C.I.40300) by a combined process of Fenton oxidation and enzymatic degradation in a sequential or mixed batch reactor is discussed. Experiments were first conducted with the enzymatic and chemical oxidation processes separately in order to determine the effects of various parameters such as pH, ferrous ion, hydrogen peroxide, dye and enzyme concentrations on the overall yield and kinetics of both processes. Decolourization was followed by UV-visible spectroscopy and high performance liquid chromatography (HPLC). Results showed that 89.5 % of DY106 were removed by enzymatic treatment after only 2 min, while 10 min of contact time were necessary to eliminate more than 98 % of 50 mg/L by Fenton’s process. A high performance was achieved under optimized conditions by the mixed combined process with time reduction down to 5 min. The study was also conducted to evaluate the efficiency of combined Fenton’s reaction as a pre-treatment and post treatment process combined with C-peroxidase at different ferrous ions concentrations. The optimal doses of Fe2+ were 2.5 mM, 1m M and 0.8 mM for Fenton, Fenton-enzymatic sequential and mixed processes. Intermediate products absorbing in UV range were detected for single Fenton or enzymatic treatment but were eliminated in all combined enzymatic-Fenton processes. Phytotoxicity tests showed that no toxicity was detected after treatment by combined process.

Celiac disease and non-celiac gluten sensitivity are both related to gluten consumption. In sensitive subjects, gluten triggers an immune-mediated enteropathy for which no therapy is yet available. Beer must be avoided by gluten sensitive subjects, since it is generally made from gluten containing grains such as barley and wheat. In this study we compared two different deglutinization treatments on an American India Pale Ale, brewed on a craft beer pilot plant: an enzymatic process based on Aspergillus niger prolyl endopeptidase and a precipitation process by silica gel. The gluten amount in all stages of brewing, and the effects of treatments on standard quality attributes, chemical composition and sensory profile were evaluated. Both treatments were able to reduce gluten level below 20 ppm, showing a different behavior. Only slight differences were observed in the chemico-physical characteristics of treated and untreated beers, whereas sensory analysis did not highlight any change. Industrial relevance Considering the massive increase of microbreweries and the grown of gluten-free products market, two different deglutinization treatment were compared in pilot plant production of craft beer. Results show how it is possible to produce a gluten-free beer from traditional ingredients. The slight differences observed between treatments had no impact on the sensory profile, allowing to brew high quality gluten-free beers.

This research is dedicated to explore practicable processes for natural bamboo fiber production. The mechanical process, steam explosion, chemical process, enzymatic process, and combined process w...

Mercury pollution is dangerous to health. Previous research was found two potential Gram-negative bacteria for mercury bioremediation, from gold mining in Central Kalimantan, Indonesia. These isolates were identified as Pseudomonas aeruginosa KHY2 and Klebsiella pneumonia KHY3. Mechanisms of mercury bioremediation had not known yet by these isolates. This study purposed to test the role of coenzymes on mercury bioremediation by these isolate and to determine the coenzymes best level of mercury bioremediation. Experimental design was Completely Randomized Design in a laboratory. Treatment factors were coenzymes obtained from vitamins B1, B6, B12, with 6 levels of treatments, included 1 control. All treatments were done in Luria Broth media that contain 12 ppm of mercury. Mercury was measured by AAS Shimadzu AA-6200. The results showed that coenzymes effect was very significant to improve mercury bioremediation by P. aeruginosa KHY2 and K. pneumonia KHY3. Supplementation of vitamin B12 in culture media, more enhance of mercury bioremediation compared with vitamin B1 and B6. These result above, indicated the mechanism of mercury bioremediation in both isolates, were the enzymatic process.

Snakehead fish (Channa striata) in South Sumatra, Indonesia has been widely used as a raw material in the processing of typical Palembang food industry, namely pempek, kemplang, and kerupuk. During its processing, not all parts of the fish can be utilized. In general, only 40% of the fish is used for consumption and 60% is wasted as by-products. One of the by-product produced during processing is viscera which can still be used as value-added products. One alternative is to use viscera as raw material in the production of fish protein hydrolysate (FPH) which is well known to have functional properties such as antioxidant, antibacterial, antihypertensive, and anticancer. This research was aim to determine the antioxidant activity of FPH made from snakehead fish viscera as a source of natural antioxidants. The research was conducted by papain treatments with the concentration of enzymes used were 1%, 2% and 3%. Evaluation of the hydrolysis process was done by measure the soluble protein and DH (degree of hydrolysis) while antioxidant activity was carried out by DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2-Azinobis 3-ethyl benzothiazoline6-sulfonic acid) assay. DH of fish protein hydrolysate was found to be 68.79%, 77.06% and 72.63%, respectively while protein solubility gave values of 5.69%, 6.15% and 6.56% respectively. The highest antioxidant activity was attained at enzyme concentration of 3% with DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2-Azinobis 3-ethyl benzothiazoline 6-sulphonic acid) value were 37.26% and 58.58% respectively.

(S)-2-Chloro-1-(3,4-difluorophenyl)ethanol (1) is a vital chiral intermediate for the synthesis of Ticagrelor—an effective treatment for acute coronary syndromes. A ketoreductase (KRED) KR-01 in our KRED library was screened to transform 2-chloro-1-(3,4-difluorophenyl)ethanone (2) into the chiral alcohol 1. During process optimization, the bioreduction procedure was performed at a substrate concentration of 500 g/L, giving a near 100% conversion with >99.9% ee. The product 1 was directly obtained by extraction and can be used for the synthesis of (1R,2R)-2-(3,4-difluorophenyl)cyclopropanecarboxylic acid ethyl ester (3) with a yield of 98% and >99.9% de, greatly simplifying the original process operation and reducing the safety risk. This process is green and environmentally sound with high productivity of biocatalysis and a space–time yield of 145.8 mmol/L/h. It has an opportunity to be very useful in industrial applications. Additional studies have indicated that KR-01 can also be used to prepare (R)-1-(...

Natural antioxidants have received considerable research interest nowadays, especially their usage in oil products. Blueberry anthocyanin extracts were added to crude camellia seed oil and lipophilized with free fatty acids in the oil through enzymatic process. The results showed that lipophilized anthocyanin derivates were synthesized, the oxidative stability of the oil under high temperature was improved, and the content of free fatty acids of the crude camellia seed oil was decreased. In conclusion, enzymatic lipophilization of blueberry extracts with free fatty acids in crude camellia seed oil could improve the quality of oil, especially the performance under high temperature.

The fate of estrogens in surface water is mainly dependent on two processes, i.e. photodegradation and biotransformation. Each of the separate process is invariably of interest, but research on the combination of the two processes has rarely been explored. In the present work, the transformation of 17α-ethinylestradiol (EE2) by simultaneous photochemical and enzymatic process in water was systematically investigated. The combined transformation rate of EE2 (0.057 h −1 ) in the presence of natural organic matter (NOM) and horseradish peroxidase (HRP) under simulated sunlight irradiation was markedly faster than that in the presence of NOM only (0.032 h −1 ). Similar pattern was also observed in real water matrix sampled from Taihu Lake. Further study revealed that the photodegradation and enzymatic transformation of EE2 were dramatically affected by NOM concentrations ranging from 0 to 20 mgC L −1 . NOM was found to invariably accelerate the photodegradation of EE2 with increasing concentration. On the contrary, the transformation rate of EE2 mediated by HRP was decreased along with the increase of NOM concentration. The reason may be that HRP was prone to be inactivated in solution with high NOM concentration. The transformation experiment of EE2 at ambient level under sunlight confirmed the significant contribution of HRP to the degradation of EE2 in the presence of NOM. The results indicated that oxidation mediated by HRP was an essential fate of EE2 and other congener contaminants in aquatic environment.

One-pot sequential asymmetric catalytic reactions as demonstrated in Table 1 represent one of the simplest kinds of enzymatic multiple-step catalytic processes. Yet quantitative analysis in these processes is more complicated than that in single-step kinetic resolutions or desymmetrization reactions, leading to lacking of a simple procedure for handling the kinetics in practical applications. Therefore based on the classification of substrate type and enzyme stereopreference for each reaction step, a thorough kinetic analysis for obtaining the analytical solution in each classification is carried out, with which the sufficient condition as an index for assessing an effective asymmetric enzymatic process is proposed. Case studies of monitoring the previous experimental data via the analytical solution and for fulfilling an assigned target for the intermediate or product are also addressed, showing the validity of the present analysis for the handling of kinetics in practical applications.

One essential engineering problem when developing an industrial enzymatic process concerns the choice of the reactor operating alternative based on a-priori knowledge of the process kinetics and enzyme inactivation characteristics. For a multi-enzymatic system, involving complex interactions among enzymes that exhibit optimal activity on different parametric domains, and a high-order deactivation, this problem requires an extended analysis. The optimization and control engineering problems become very challenging due to the process high nonlinearity and the presence of a large number of complex constraints. All these will translate into a multi-objective optimization problem to be solved for each case. An elegant option developed in this paper is to obtain sets of Pareto optimal solutions, also called Pareto-optimal fronts, successively generated for pairs of adverse objectives. Then, the final choice of the enzymatic reactor operating policy results from the comparative analysis of these Pareto-fronts. Exemplification is made for the complex case of the oxidation of D-glucose (DG) to 2-keto- d -glucose (kDG) in the presence of P2Ox (Pyranose oxidase, EC 1.1.3.10) and catalase, continuously operated in a three-phase mechanically agitated semibatch reactor (MASCR) with co-immobilized enzymes on alginate beads. Optimal operating policy choice is based on the minimum amount of required P2Ox that ensures an imposed reaction conversion and maximum reactor productivity under various technological constraints, and for catalase/P2Ox ratios of 200–300 U/U.

Ansa Biotechnologies has raised $7.9 million in seed financing, bringing its funding to date to $9.2 million. The California-based company synthesizes DNA with a fully enzymatic process that it cla...

Nanotechnology has leveraged the Industrial Biotechnology’s processes and products by inclusion of nanoscale materials in the bioprocessing application to make cost-effective recyclable biocatalyst at the industrial setting. Conventional recyclable enzymes, based on bulk supports, have been used in traditional industries; however, they have limitations of mass transfer and low enzyme loading that make enzymatic process unfavorable and inefficient for commercial applications. With the intervention of functional nanoscale material in the range of 1–100 nm as a nanocarrier for industrial biocatalysts, enzymatic process becomes attractive for higher productivity due to higher enzyme loading, faster mass-transfer rate, and ease of bioseparation. The present chapter is focused on the nanobiocatalyst design and subsequent application in food industry, with reference to concentrating omega-3 fatty acid.

Cellobiose is a naturally occurring only slightly sweet disaccharide that may be used as a novel food ingredient. As part of the safety evaluation of cellobiose, produced from sucrose by an enzymatic process, an Ames test and an in-vitro micronucleus test with human peripheral lymphocytes were performed according to the respective OECD Guidelines. The results demonstrate that cellobiose lacks a genotoxic potential under the conditions of these standardized assays.

Anticancer drugs are a class of pharmaceutical compounds that have been found in hospital, domestic, and industrial wastewaters and also in surface waters. They have been showing recalcitrance to conventional wastewater treatment technologies and present a potential risk to environment and human health, since they exhibit cytotoxic, teratogenic, and carcinogenic among other effects in higher organisms, even at low concentrations. The presence of these compounds in the environment is a recent challenge for wastewater treatment and some alternative strategies to remove them were already studied, such as white-rot fungi (WRF) technologies. Despite promising results, processes involving fungi are complex, have high reaction times, and require nutrient addition for fungus growth and maintenance. Due to this potential, strategies to make the technology feasible were studied, such as the possibility for direct application of enzymes secreted by WRF. Enzymatic processes were studied in the removal of other pharmaceuticals such as antibiotics, anti-inflammatory, and steroid hormones; however, to the best of our knowledge, there is a gap on literature about their direct action on anticancer drugs.

Background: Stromal vascular fraction cells (SVFs), which can be produced using a mechanical or digestive enzymatic process, are heterogeneous cells with the potential to grow hair in androgenic alopecia patients. Platelet-rich plasma (PRP) is an autologous cell source that is widely used to treat androgenic alopecia. However, the combined use of PRP and SVFs to treat alopecia is rarely reported. Case presentation: This case series describes three cases of androgenic alopecia, including a32-year-old male, a 43-year-old male, and a 65-year-old male. Androgenic alopecia in these patients was treated by injecting the bald area of the scalp with local autologous PRP combined with SVFs. Follow-up was performed 1–3 months after treatment, which showed good results. The hair grew denser, with increased thickness for each strand. The hair pull test revealed that the hair remained intact. Conclusion: The combination of PRP and SVFs affected all cycles of the hair growth process. SVFs are multipotent cells with the potential to become antioxidant, anti-inflammatory, or anti-fibrotic cells. SVFs can regenerate cells that secrete the growth factors, that are essential for angiogenesis, which can improve therapeutic outcomes. This case series will enrich the existing literature by expanding available treatment options for androgenic alopecia.

Activation of O–H bonds by inorganic metal-oxo complexes has been documented, but no cognate enzymatic process is known. Our mechanistic analysis of 2-hydroxyethylphosphonate dioxygenase (HEPD), which cleaves the C1–C2 bond of its substrate to afford hydroxymethylphosphonate on the biosynthetic pathway to the commercial herbicide phosphinothricin, uncovered an example of such an O–H-bond-cleavage event. Stopped-flow UV–visible absorption and freeze-quench Mössbauer experiments identified a transient iron(IV)-oxo (ferryl) complex. Maximal accumulation of the intermediate required both the presence of deuterium in the substrate and, importantly, the use of 2H2O as solvent. The ferryl complex forms and decays rapidly enough to be on the catalytic pathway. To account for these unanticipated results, a new mechanism that involves activation of an O–H bond by the ferryl complex is proposed. This mechanism accommodates all available data on the HEPD reaction.

In this paper, immobilized laccase enzyme on nano zinc ferrite was used in order to decolorize disperse dyes from single and binary systems. In this case, disperse dyes such as Disperse red 60 (DR60), Disperse blue 56 (DB56) and Disperse yellow 54 (DY54) were selected as model dyes. Several parameters such as enzyme concentration, pH and dye concentration and their effect on decolorization of dyes from single and binary systems were studied. According to the experimental results, the optimized immobilized laccase enzyme concentration, reaction time and pH for decolorization of DR60, DB56 and DY54 from single and binary systems were 500 mg/L (for DR60 and DY54) and 400 mg/L (for DB56), 20 min and 3, respectively. Moreover, Dye decolorization kinetics followed Michaelis-Menten Model. Finally, the results showed that enzymatic process using immobilized laccase enzyme on nano zinc ferrite was effective method to decolorize disperse dyes from single and binary systems.

Deamidation of glutamine (Gln) residues is a spontaneous or enzymatic process with significant implications in aging and human pathology. Although some methods are available to identify the γ/α-glutamyl products of deamidation, none of these methods allows the characterization of this post-translational modification (PTM) from complex biological samples by shotgun proteomics. Here we present LERLIC-MS/MS, a chromatographic strategy that uses a long (50 cm) anion-exchange capillary column operating in the electrostatic repulsion-hydrophilic interaction mode (ERLIC) and coupled directly to tandem mass spectrometry (MS/MS) for proteome analysis in a single injection. Profiling of soluble extracts of brain tissues by LERLIC-MS/MS distinguished for the first time γ/α-glutamyl isomers of deamidation, encountering a 1.7 γ/α-glutamyl ratio for most Gln deamidation products. A detailed analysis of any deviation from that observed ratio allowed the identification of transglutaminase-mediated γ-glutamyl isomers as intermediate products of transamidation. Furthermore, LERLIC-MS/MS was able to simultaneously separate Gln and asparagine (Asn) deamidation products even for those peptides showing multiple deamidated proteoforms. The characterization of Asn deamidated residues by LERLIC-MS/MS also uncovered novel PIMT (protein L-isoaspartyl methyltransferase) substrate proteins in human brain tissues that deviated from the expected 3:1 isoAsp/Asp ratio. Taken together, our results demonstrate that LERLIC-MS/MS can be used to perform an in-depth study of protein deamidation on a global proteome scale. This new strategy should help to elucidate the biological implications of deamidation in aging and disease conditions. MOE (Min. of Education, S’pore) Published version