BIOTRANSFORMATION OF 2α，5α，10β，14β－TETRA－ACETOXY－4（20），11－TAXADIENE BY THE FUNGIJS CUNNINGHAMELLA ECHINULATA

?5α,10β,14β-Tetra-acctoxy-4(20).11-taxadicne (1) could be regioselectively transfonnedinto 6α 10β-dihydroxy-2α,5α,14β-triacetoxy-4(20). 11- taxadiene (2) by Cunninghamella echinnlata infair yield. 6βM.10β-Dihydroxy-2α,5α,14β-triacet 11-taxadienc (3). 10β-hydroxy-2α,5α,14β-triacetoxy-4(20).11-taxadiene (4), 10 β-hyboxy-4β,20-epoxy -2α,5α,14β-triacetoxy- tax - 11- ene (5)asminor prpducts were also isolated

Biotransformation of 14-Deacetoxy-13-oxo sinenxan A by Ginkgo Cell Cultures

Deacetoxy-13-oxo sinenxan A (1) was converted to 9a-hydroxy-13-oxo-2a, 5a, 10b-triacetoxy-4(20),11-taxadiene (2) and 10b-hydroxy-13-oxo-2a,5a,9a-triacetoxy- 4(20), 11- taxadiene (3) by Ginkgo cell suspension cultures in 45% and 15% yields, respectively.

Time-course monitoring of in vitro biotransformation reaction via solid-phase microextraction-ambient mass spectrometry approaches

The solid-phase microextraction technique quantifies analytes without considerably affecting the sample composition.Herein,a proof-of-concept study was conducted to demonstrate the use of coated probe electrospray ionization(coated-PESI)and coated blade spray(CBS)as ambient mass spectrometry approaches for monitoring drug biotransformation.The ability of these methods was investigated for monitoring the dephosphorylation of a prodrug,combretastatin A4 phosphate(CA4P),into its active form,combretastatin A4(CA4),in a cell culture medium supplemented with fetal bovine serum.The CBS spot analysis was modified to achieve the same extraction efficiency as protein precipitation and obtained results in 7 min.Because coated-PESI performs extraction without consuming any samples,it is the preferred technique in the case of a limited sample volume.Although coated-PESI only extracts small quantities of analytes,it uses the desorption solvent volume of 5-10 pL,resulting in high sensitivity,thus allowing the detection of compounds after only 1 min of extraction.The biotransformation of CA4P into CA4 via phosphatases occurs within the simple matrix,and the proposed sample preparation techniques are suitable for monitoring the biotransformation.

Fatty acid hydratase for value-added biotransformation:A review

The synthesis of hydroxy fatty acids(HFAs) from renewable oil feedstock by addition of water onto C_C bonds has attracted great attention in recent years. Given that selective asymmetric hydration of non-activated C_C bonds has been proven difficult to achieve with chemical catalysts, enzymatic catalysis by fatty acid hydratases(FAHs) presents an attractive alternative approach to produce value-added HFAs with high regio-, enantioand stereospecificity, as well as excellent atom economy. Even though FAHs have just been investigated as a potential biocatalyst for a decade, remarkable information about FAHs in different aspects is available;however, a comprehensive review has not been archived. Herein, we summarize the research progresses on biochemical characterization, structural and mechanistic determination, enzyme engineering, as well as biotechnological application of FAHs. The current challenges and opportunities for an efficient utilization of FAHs in organic synthesis and industrial applications are critically discussed.

Herba Epimedii′s biotransformation in vitro simulated gastrointestinal digestion and faecal fermentation systems

OBJECTIVE Epimedium is rich in a variety of beneficial active ingredients,and has been widely used in the ethnopharmacological practices,however,its biotransformation in gastrointestinal digestions remain unclear.This study aimed to investigate the dynamic changes of components and biological activity of Epimedium in the in vitro simulated digestion and subsequent human faecal fermentation.METHODS The models of in vitro simulated saliva,gastric and intestinal digestion,as well as colonic fermentation were constructed to simulate the digestion process of Epimedium.The dynamic changes of components of Epimedium during the simulated digestions in vitro and subsequent human faecal fermentation were investigated by UPLC-MS,HPLC-DAD combined with principal component analysis(PCA)and multi-ingredient quantitative analysis.RESULTS A variety of metabolites with high contents were produced after 0.5 h of intestinal digestion and colonic fermentation 0.5 h.Application of PCA to HPLC data showed the obvious separation of colonic fermentation 0.5 h stage samples from other colonic fermentation stages samples(24,48 and 72 h).Additionally,non-digestion and saliva digestion stage samples clustered together,and there was obvious separation between intestinal digestion samples and gastric digestion samples.The contents of epimedium C,icariin and baohuside I all increased significantly after intestinal digestion[58.70±7.08,47.15±5.68 and(12.78±0.55)mg·g^(-1)]compared with gastric digestion[29.00±5.65,17.40±4.55 and(2.77±0.19)mg·g^(-1)].There were significant differences between sample after 0.5 h of colonic fermentation[64.22±9.32,51.26±6.33 and(16.68±3.19)mg·g^(-1)]and other time points(24,48 and 72 h)in components and the contents of active ingredient,and the content of these components all decreased with the fermentation time.The ability of scavenging ABTS free radicals[IC50=(0.29±0.02)g·L^(-1)]increased significantly compared with gastric digestion[(1.57±0.02)g·L^(-1)],and after 0.5 h of colonic fermentation,the ability also increased significantly.CONCLUSION Gastrointestinal digestion had a significant impact on the contents of active components in Epimedium,and the metabolism of these components mainly occurred in the colon.The intestinal digestion and colonic fermentation significantly improved the anti-ABTS activity of epimedium.

Biotransformation of Flavor Compositions During Fermentation of Litchi (Litchi Chinensis Sonn.) Fruits into Wine

The aim was to examine the biotransformation of chemical compounds during the fermentation of litchi wine.S.bccyanus BV818 was inoculated to litchi juice(Heiye)to initiate the fermentation.Acetic acid decreased dramatically,succinic acid and DL-malic acid increased sharply.Saturated free fatty acid increased,especially the concentration of the free fatty acid with long carbon chain(more than 10 carbons)increased significantly.The unique flavor compounds of fresh litchi including linalool,α-terpineol,β-citronellol and other terpenoids remained in the litchi wine were transformed to other aroma constituents,by which the primary litchi flavor was retained.The wine had a fruity flavor and delicate bouquet and had harmonious sourness and sweetness.The litchi'Heiye'was suitable for being fermented into litchi wine.

Biotransformation of Carmoisine and Reactive Black 5 Dyes Using <i>Saccharomyces cerevisiae</i>

Saccharomyces cerevisiae (baker’s yeast) is the most important industrial microorganisms. This yeast is commonly used as a leavening agent in baking bread and bakery products, where it produces carbon dioxide from converting of the fermentable sugars present in the dough. Nowadays, industrial and chemical activities led to produce new compounds with new kinds of contamination in the environment. Discharge of untreated or partially treated industrial sewage has created the contamination problems of rivers and lakes such as drugs, oil, heavy metals, paints, pesticides and various chemical compounds in them. Hence, it is necessary to control and reduce the levels of these compounds in wastewater and bring them to permissible values. This study aims to study the bioconversion potential of commonly available Saccharomyces cerevisiae for the two textile dyes of Carmoisine and Reactive Black 5. Reaction mixtures for biotransformation of dyes included 50 mg/l Carmoisine or 25 mg/l Reactive Black 5 and 1% dried harvested cells of S. cerevisiae (bread’s yeast) were tested. Harvested dry and wet yeast were studied for this purpose. The results show that harvested cells of Saccharomyces cerevisiae are able to bioconvert Carmoisine and Reactive Black 5. Reactive Black 5, Carmoisine are degraded by biotransformation 85% and 53% within 24 hours in water at the room temperature.

Unbalanced biotransformation metabolism and oxidative stress status: implications for deficient fatty acid oxidation

The concept of accumulating xenobiotics within the human body as a health risk is well known. However, these compounds can also be endo-genous, as in the case of inborn errors of me-tabolism, and lead to some of the same symp-toms as seen in xenobiotic intoxication. Bio-transformation of both exogenous and endo-genous toxic compounds is an important function of the liver, and the critical balance between these systems is of fundamental importance for cellular health. We propose a novel model, to describe the critical balance between Phase I and Phase II biotransformation and how a disturbance in this balance will increase the oxidative stress status, with resulting pathological consequences. We further used deficient fatty acid oxidation to verify the proposed model, as deficient fatty acid oxidation is associated with the accumulation of characteristic metabolites. These accumulating metabolites undergo both Phase I and Phase II biotransformation reactions, with resulting depletion of biotransformation substrates and co-factors. Depletion of these important biomolecules is capable of disturbing the balance between Phase I and Phase II reactions, and disturbance of this balance will increase oxidative stress status. The value of the proposed model is illustrated by its application to a clinical case investigated in our laboratory. In this case the possibility of deficient fatty acid oxidation only became evident once the critical balance between Phase I and Phase II biotransformation was restored with oral replenishment of biotransformation substrates. In addition to bio-chemical improvement, there was also significant clinical improvement. The significance of this model lies within the treatment possibilities, as the assessment of biotransformation metabolism and oxidative stress status can lead to the development of nutritional treatment strategies to correct imbalances. This in turn may reduce the chances of, or delay the onset of certain disease states.

Biotransformation of 6-deoxypseudoanisatin by Synechocystis sp. PCC6803

Objective:To explore the ability of Synechocystis sp.PCC6803 in transforming 6-deoxypseudoanisatin.Methods:The experiment was performed by incubating 6-deoxypseudoanisatin with the freshwater cyanobacterium Synechocystis sp.PCC6803 under continuous white light at 30C for 5 days.The crude converted product was detected using thin-layer chromatography(TLC)and further analyzed using high-performance liquid chromatography(HPLC)as well as HPLC with electron spray ionization mass spectrometry(HPLC-ESI-MS).Results:TLC results showed that 6-deoxypseudoanisatin was converted into a less polar product.HPLC and MS data indicated that the retention time of the converted product increased in comparison with the standard of 6-deoxypseudoanisatin.Conclusion:Thus,the study appears to demonstrate that Synechocystis sp.PCC6803 can transform 6-deoxypseudoanisatin.The polarity of the converted product is less than that of 6-deoxypseudoanisatin.

Study on Biotransformation of Natural Anti-tumor Drug Podophyllotoxin

[Objective] This study aimed to select stains for transforming podophyllotoxin, to isolate the transformation product and to identify the structure. [ Method~ Structure of podophyllotoxin was modified with microbial batch transformation method. Podophyllotoxin and its transformation products were detected by using TLC and HPLC methods to select strains with ability to transform podophyllotoxin. Scale-up fermentation of one podophyllotoxin-transforming strain was carried out and the fermented material was extracted with EtoAc, isolated with conventional silica gel column chromatography, structure of the transformation product was identified by using classic spectroscopy technology. [ Result ] Based on researches of pedophyllotoxin transformation, seven strains with ability to transform po- dophyllotoxin were selected, including BS. Str-1, BS. Str-2, BS. Str-3, BS. Str-7, BS. Str-18, BS. Str-21 and BS. Str-29. By scale up fermentation, the only trans- formation product of BS. Str-21 strain was extracted and isolated, which was identified as picropodophyllotoxin. [ Conclusion] This study demonstrates that podo- phyllotoxin derivatives can be obtained by biotransformation methods, which lays the foundation for acquisition of natural drugs with high anti-tumor activity.

Anaerobic Biotransformation of Nitro-Compounds to Amines by Bovine Rumen Fluid

Microorganims of the bovine rumen fluid biocatalyzed the reduction of nitro-compound substrates to yield the respective amines. This enzymatic process, using ruminal contents, has rarely been reported in associa- tion with the bioreduction of nitro groups. The biotransformation reactions catalyzed by this system were de- pendent of both the electronic characteristics and the area/volume of the nitro-substrates confirming the processes are enzymatic. The semi-preparative scale biotransformation went by in good yield showing the rumen fluid may be employed in the synthesis of amines under very mild conditions and, moreover, it may have application in the bioremediation of nitro-compounds.

Production of Enzymes and Biotransformation of Orange Waste by Oyster Mushroom, <i>Pleurotus pulmonarius</i>(Fr.) Quél.

The wood-decay fungi are able to bioconvert a wide variety of lignocellulosic residues due to the secretion of extracellular enzymes. The use of agricultural wastes as substrate for mushroom cultivation or enzymes production can help to solve environmental problems caused by inadequate discharge in the nature. The production of hydrolytic and oxidative enzymes by Pleurotus pulmonarius developed in solid state system using orange waste as substrate was evaluated in this work. Among the hydrolytic enzymes, pectinase was the main enzyme produced by the fungus, presenting the highest enzymatic activity of 9.4 U/mL after 35 days of cultivation. Considering the oxidative enzymes, laccase was the main enzyme produced with maximal activity of 12.2 U/mL obtained after 20 days of cultivation. Low enzyme levels of manganese peroxidase, β-glucosidase and β-xy-losidase were detected with activity peaks at the end of the cultivation. The enzymatic levels of amylase, carboxymethyl cellulase and xylanase were similar and less than 1.5 U/mL. No aryl-alcohol oxidase activity was detected. NDF, ADF and cellulose values increased during 45 days of cultivation. There was no lignin degradation during the study period and the fungus culture in orange solid waste caused protein enrichment in the substrate. Our results demonstrate that P. pulmonarius was an efficient producer of two important industrial enzymes, pectinase and laccase in a cheap solid state system using orange waste as substrate.

Study on Biotransformation of Traditional Chinese Herbal Medicines Complex

A newly designed enzymolysis-fermentation combined method to dramatically enhance the actives level and skin benefits of traditional Chinese herbs (TCHs) was developed, biotransformation process under optimal reaction conditions can significantly transform molecular structures and obtained fermentation extracts can deliver better skin benefits on anti-aging, hydration and whitening. Analytical results showed that the ginsenoside Rg3, total sugar, polyphenols and flavonoids in fermented extract were 2~4 times higher than unfermented extract. In-vitro tests including DPPH radical scavenging activity, tyrosinase inhibition, cells proliferation and Hyaluronan-CD44 activity, have showed significant enhancement of efficacy, inhibition rate on DPPH antioxidation as example achieved over 60% improvement, which makes traditional Chinese herbs application more feasible. It is inferred that this study potentially enables the herbs to deliver required bioefficacy in cosmetic application.

Mimotope peptide modified pompon mum-like magnetic microparticles for precise recognition,capture and biotransformation analysis of rituximab in biological fluids

Due to low immobilized ligand density,limited binding capacity,and severe interference from serum proteins,developing ideal peptide-based biomaterials for precise recognition and in vivo analysis of biopharmaceuticals remains a huge challenge.In this study,mimotope peptide modified pompon mum-like biomimetic magnetic microparticles(MMPs,3.8μm)that mimic the specific functionalities of CD20 on malignant B cells were developed for the first time.Benefit from the numerous ligand binding sites(Ni^(2+))on the pompon mum-like MMPs,these novel materials achieved≥10 times higher peptide ligand densities(>2300 mg/g)and antibody binding capacities(1380 mg/g)compared to previous reported biomaterials.Leveraging the high specificity of the mimotope peptide,rituximab can be precisely recognized and enriched from cell culture media or serum samples.We also established an LC-MS/MS method using the MMPs for tracking rituximab biotransformation in patient serum.Intriguingly,deamidation of Asn55 and Asn33,as well as oxidation of Met81 and Met34 were observed at the key complementarity determining regions of rituximab,which could potentially influence antibody function and require careful monitoring.Overall,these versatile biomimetic MMPs demonstrate superior recognition and enrichment capabilities for target antibodies,offering interesting possibilities for biotransformation analysis of biopharmaceuticals in patient serum.

Co-occurrence of genes for antibiotic resistance and arsenic biotransformation in paddy soils

Paddy soils are potential hotspots of combined contamination with arsenic(As) and antibiotics, which may induce co-selection of antibiotic resistance genes(ARGs) and As biotransformation genes (ABGs), resulting in dissemination of antimicrobial resistance and modification in As biogeochemical cycling. So far, little information is available for these coselection processes and specific patterns between ABGs and ARGs in paddy soils. Here, the16S rRNA amplicon sequencing and high-throughput quantitative PCR and network analysis were employed to investigate the dynamic response of ABGs and ARGs to As stress and manure application. The results showed that As stress increased the abundance of ARGs and mobile genetic elements (MGEs), resulting in dissemination risk of antimicrobial resistance. Manure amendment increased the abundance of ABGs, enhanced As mobilization and methylation in paddy soil, posing risk to food safety. The frequency of the co-occurrence between ABGs and ARGs, the host bacteria carrying both ARGs and ABGs were increased by As or manure treatment, and remarkably boosted in soils amended with both As and manure. Multidrug resistance genes were found to have the preference to be co-selected with ABGs, which was one of the dominant co-occurring ARGs in all treatments, and manure amendment increased the frequency of Macrolide-Lincosamide-Streptogramin B resistance(MLSB) to co-occur with ABGs. Bacillus and Clostridium of Firmicutes are the dominant host bacteria carrying both ABGs and ARGs in paddy soils. This study would extend our understanding on the co-selection between genes for antibiotics and metals, also unveil the hidden environmental effects of combined pollution.

Biotransformation,multifunctional recycling mechanism of nanostructures,and evaluation of the safety of nanoscale materials

Current evidence of concept analyses recommending nanotechnology for biomedical uses abounds in recent research. The area of biotechnology interfaces with nanostructures, reconfigures their composition, and alters their characteristics;which influences the dispersion of the particles, the biotransformation they cause, and their potential toxic effect. It is vital to link the idea of the lifecycle of nanostructures to the biological impacts and use methodologies to identify, estimate, and track the gradual bioprocessing of nanostructures in vivo, from a body-wide level to a nanoscopic size. This is necessary because understanding how nanostructures processing, degradation, persistence, and recycling predict potential exposure risks. The safe implementation of nanotechnology-based products in biomedical applications necessitates an extensive understanding of the recycling and transformations of nanomaterials in a living organism. Long-term fate in the body is crucial, as it governs potential environmental risks to human health. Strategies may be used to manage the long-term outcome of nanostructures in an organism since, in addition to composition, their design also affects how long they last and how easily they degrade. The lifespan of nanoparticles, a flexible and biocompatible category of nanostructures that have made it into clinical trials, is the subject of this article. Strategies may be used to manage the long-term outcome of nanoparticles in an organism since, in addition to composition, their design also affects how long they last and how easily they degrade. This review explained the safety of nanoscale materials, biotransformation, and the multifunctional recycling mechanism of nanostructures.

Biotransformation differences of ginsenoside compound K mediated by the gut microbiota from diabetic patients and healthy subjects

Many natural products can be bio-converted by the gut microbiota to influence pertinent efficiency.Ginsenoside compound K(GCK)is a potential anti-type 2 diabetes(T2D)saponin,which is mainly bio-transformed into protopanaxadiol(PPD)by the gut microbiota.Studies have shown that the gut microbiota between diabetic patients and healthy subjects are significantly different.Herein,we aimed to characterize the biotransformation of GCK mediated by the gut microbiota from diabetic patients and healthy subjects.Based on 16S rRNA gene sequencing,the results indicated the bacterial profiles were considerably different between the two groups,especially Alistipes and Parabacteroides that increased in healthy subjects.The quantitative analysis of GCK and PPD showed that gut microbiota from the diabetic patients metabolized GCK slower than healthy subjects through liquid chromatography tandem mass spectrometry(LC-MS/MS).The selected strain A.finegoldi and P.merdae exhibited a different metabolic capability of GCK.In conclusion,the different biotransformation capacity for GCK may impact its anti-diabetic potency.

The dynamic changes of arsenic biotransformation and bioaccumulation in muscle of freshwater food fish crucian carp during chronic dietborne exposure

Dietary uptake is the major way that inorganic arsenic(iAs)enters into benthic fish;however,the metabolic process of dietborne i As in fish muscle following chronic exposure remains unclear.This was a 40-day study on chronic dietborne i As[arsenite(AsⅢ)and arsenate(AsⅤ)]exposure in the benthic freshwater food fish,the crucian carp(Carassius auratus),which determined the temporal profiles of iAs metabolism and toxicokinetics during exposure.We found that an adaptive response occurred in the fish body after iAs dietary exposure,which was associated with decreased As accumulation and increased As transformation into a non-toxic As form(arsenobetaine).The bioavailability of dietary AsⅢwas lower than that of AsⅤ,probably because AsⅢhas a lower ability to pass through fish tissues.Dietary AsⅤexhibited a high potential for transformation into AsⅢspecies,which then accumulated in fish muscle.The largely produced AsⅢconsidered more toxic at the earlier stage of AsⅤexposure should attract sufficient attention to human exposure assessment.Therefore,the pristine As species and exposure duration had significant effects on As bioaccumulation and biotransformation in fish.The behavior determined for dietborne arsenic in food fish is crucial for not only arsenic ecotoxicology but also food safety.

Biotransformation of 4-Hydroxybenzen Derivatives by Hairy Root Cultures of Polygonum multiflorum Thunb

The biotransformation of four 4-hydroxybenzen derivatives （1,4-benzenediol （compound 1）, 4-hydroxybenzaldehyde （compound 2）, 4-hydroxybenzyl alcohol （compound 3） and 4-hydroxybenzoic acid （compound 4）） by the hairy root cultures of Polygonum multiflorum Thunb. as a new biocatalyst was investigated. It was found that the substrates were transformed to their corresponding glucosides, 4-hydroxyphenyl β-D-glucopyranoside （arbutin, compound la）, 4-hydroxymethylphenyl β-D-glucopyranoside （gastrodin, compounds 2a, 3a） and 4-carboxyphenyl α-D- glucopyranoside （compound 4a）, respectively. In the meantime, the hairy roots of P. multiflorum were able to stereoselectively and regioselectively glucosylate phenolic hydroxyl groups of compounds 1-4, but the cultures could not glucosylate the aldehyde group of compound 2 or the benzyllc hydroxyl group of compound 3, and no glucosyl esterification of carboxyl groups of compound 4 was detected. On the other hand, the result also showed that the hairy roots of P. multiflorum were able to reduce the 4-hydroxybenzaldehyde to its corresponding alcohol. This is the first report that substrate 4 has been converted into its α-D-glucopyranoside by a plant biotransformα- tion system.