Advances in biotransformation of methanol into chemicals / 生物工程学报

Methanol has become an attractive substrate for the biomanufacturing industry due to its abundant supply and low cost. The biotransformation of methanol to value-added chemicals using microbial cell factories has the advantages of green process, mild conditions and diversified products. These advantages may expand the product chain based on methanol and alleviate the current problem of biomanufacturing, which is competing with people for food. Elucidating the pathways involving methanol oxidation, formaldehyde assimilation and dissimilation in different natural methylotrophs is essential for subsequent genetic engineering modification, and is more conducive to the construction of novel non-natural methylotrophs. This review discusses the current status of research on methanol metabolic pathways in methylotrophs, and presents recent advances and challenges in natural and synthetic methylotrophs and their applications in methanol bioconversion.

Biosynthesis of steroidal intermediates using Mycobacteria: a review / 生物工程学报

Steroids are a class of medicines with important physiological and pharmacological effects. In pharmaceutical industry, steroidal intermediates are mainly prepared through Mycobacteria transformation, and then modified chemically or enzymatically into advanced steroidal compounds. Compared with the "diosgenin-dienolone" route, Mycobacteria transformation has the advantages of abundant raw materials, cost-effective, short reaction route, high yield and environmental friendliness. Based on genomics and metabolomics, the key enzymes in the phytosterol degradation pathway of Mycobacteria and their catalytic mechanisms are further revealed, which makes it possible for Mycobacteria to be used as chassis cells. This review summarizes the progress in the discovery of steroid-converting enzymes from different species, the modification of Mycobacteria genes and the overexpression of heterologous genes, and the optimization and modification of Mycobacteria as chassis cells.

F-box motif encoding genes as targets for the development of stress-tolerant traits in Saccharomyces cerevisiae

Stress tolerance is a useful trait actively sought by the bioprocess industry for biofactories dealing with bioconversion of varied raw materials or carbon sources. Fungal or yeast species are useful in bioconversion and are sustainable bioresources for biochemicals and biofuel production. Genetic manipulation strategies are in practice to enhance the tolerance against stress agents for the improved bioconversion process. In this review, we highlight the importance of the F-box motif encoding genes and their interactions in imparting the stress tolerance phenotype to the yeast species. The F-box motif proteins constitute a part of the SCF-E3 ligase complex and are involved in the recruitment, and ubiquitination, followed by degradation of the substrate proteins by the 26S proteasome. It highlights the current scenario on the F-box motif encoding genes and their interaction partners as targets for the stress tolerance phenotype in the yeast and plant species and their utility in the bioconversion processes.

Cytotoxicity and the bioconversion strategy of Aristolochia spp

Aristolochia plants are notable from an ethnopharmacological viewpoint, but the relevance of these species for medicinal purposes has been debated because of their inherent toxicity. The convergence of these contrasting realities can be readily achieved using bioconversion methods, which have been shown to be useful tools for numerous applications, including the detoxification of biomass. In this context, methanolic extracts of leaves from Aristolochia triangularis and Aristolochia gibertii, as well as the feces of Battus polydamas larvae fed with leaves from these plants, were prepared, and their cytotoxic activities were evaluated on a human fibroblast cell line (GM07492). The leaf extracts were found to be cytotoxic, leading to reductions of 42.1 and 33.8% on cell viability, respectively, while the fecal extracts were considered inactive. In addition to evidencing the cytotoxicity of A. triangularis and A. gibertii, these findings demonstrated a potential bioconversion strategy for obtaining aristolochiaceous extracts with reduced toxicity using the larvae of a specialist phytophagous insect, thus renewing expectations in relation to the pharmacological importance of Aristolochia spp. The results were also ecologically relevant, as B. polydamas larvae were found to be able to detoxify compounds from host plants.(AU)

Tecnologic Development on Pleurotus Cultivation: Specific Practices Used in Brazil

HIGHLIGHTS P. ostreatus and P. sapidus are the most productive species under the evaluated conditions. Different growing systems are suitable for the production of P. ostreatus var. Florida. Temperature control level affects differently the P. ostreatus var. Florida isolates. Environmental and strain factors affect yield and production parameters of P. ostreatus var. Florida.

Abstract In Brazil, Pleurotus is the most important mushroom produced especially P. ostreatus var. Florida. In this country as in many others, the great potential for mushroom cultivation remains unexplored. Therefore, it is very important to develop new studies that allow optimizing its production. The aims of the manuscript were: i) to evaluate the productivity of six different species of Pleurotus (P. citrinopileatus; P. djamor; P. ostreatus; P. ostreatus var. Florida; P. pulmonarius; P. sapidus); ii) to measure the effect of three different environmental conditions during cultivation of three isolates of P. ostreatus var. Florida. As results, P. ostreatus and P. sapidus were the most productive isolates under the evaluated conditions. Different environments produced variable effects according to the P. ostreatus var. Florida isolates, being possible to observe a highly plastic strain (POF 02/18), a highly sensitive strain (POF 03/18) and a strain with variable responses (POF 01/18).

Construction of acetyl-CoA and DBAT hybrid metabolic pathway for acetylation of 10-deacetylbaccatin III to baccatin III

10-Deacetylbaccatin III (10-DAB) C10 acetylation is an indispensable procedure for Taxol semi-synthesis, which often requires harsh conditions. 10-Deacetylbaccatin III-10-

Advances in lignin valorization from a biorefinery concept / 生物工程学报

Lignin valorization for fuels and value-added products is essential to enhance the profitability and sustainability of biorefineries. Due to the complex and heterogeneous structure of lignin, technical barriers hinder the implementation of economic lignin utilization. Here, we summarize the major challenges facing lignin valorization processes. Different pretreatment methods, especially emerging combinatorial pretreatment approaches for isolating and tailoring lignin are introduced. To overcome the heterogeneity of lignin structure and improve lignin processability, advances in fractionation approaches including organosolv extraction, membrane technology, and gradient precipitation are analyzed and presented. Furthermore, progress in lignin valorization by thermochemical and biological conversion coupling with pretreatment and fractionation are systematically reviewed. Finally, we discuss advanced strategies and perspectives for future research involving biomass pretreatment, lignin fractionation and conversion processes.

Bioconversion of methane by metabolically engineered methanotrophs / 生物工程学报

Due to abundant availability of shale gas and biogas, methane has been considered as one of the most potential carbon sources for industrial biotechnology. Methanotrophs carrying the native methane monooxygenase are capable of using methane as a sole energy and carbon source, which provides a novel strategy for reducing greenhouse gas emission and substituting edible substrates used in bioconversion processes. With the rapid development of genetic engineering tools and biosynthesis techniques, various strategies for improving the efficiency of methane bioconversion have been achieved to produce a variety of commodity bio-based products. Herein, we summarize several important aspects related with methane utilization and metabolic engineering of methanotrophs, including the modification of methane oxidation pathways, the construction of efficient cell factories, and biosynthesis of chemicals and fuels. Finally, the prospects and challenges of the future development of methane bioconversion are also discussed.

Biological efficiency and nutritional value of Pleurotus ostreatus cultivated in agroindustrial wastes of palm oil fruits and cocoa almonds / Eficiência biológica e valor nutricional de Pleurotus ostreatus cultivados em resíduos agroindustriais de dendê e amêndoa de cacau

The cocoa and palm oil agro-industries active in the state of Bahia, Brazil, generate high quantities of lignocellulosic wastes that could be recycled through their use in the formulation of substrates to cultivate edible mushrooms. Pleurotus ostreatus, also known as oyster mushroom, is the second most cultivated mushroom in the world due to its highly appreciated gastronomic, nutritional, and medicinal characteristics. This work evaluated the vertical mycelium growth, biological efficiency, mushroom yield, and nutritional composition of P. ostreatus produced in substrates formulated with a combination of palm oil fruit mesocarp (POFM) and cocoa almond peels (CAP) processing wastes. The substrates were formulated with the following POFM/CAP proportions (%/%): S1 ­ 86.4/9.6; S2 ­ 76.8/19.2; S3 ­ 67.2/28.8; S4 ­ 57.6/38.4, and S5 ­ 48.0/48.0. Substrates also received 3% powdered charcoal and 1% calcium carbonate. Substrates S1, S2, S3, and S4 were superior for vertical mycelium growth. S2 promoted the best biological efficiency (148.8%) and yield (560.5g·kg-1).The mushrooms produced in all substrates presented good nutritional values, although mushrooms produced using the S2 presented the highest crude protein content. Overall, S1 is the recommended substrate as it results in higher yields of nutrient rich mushrooms. Production of P. ostreatus in substrates composed of POFM and CAP represents a good alternative for recycling these wastes with potential economic and ecological benefits to regions where palm oil and cocoa are grown.(AU)

As indústrias de cacau e óleo de dendê no estado da Bahia, Brasil, geram grandes quantidades de resíduos lignocelulósicos que podem ser reciclados na formulação de substratos para o cultivo de cogumelos comestíveis. Pleurotus ostreatus ou cogumelo ostra é o segundo cogumelo mais cultivado no mundo por apresentar características gastronômicas, nutricionais e medicinais muito apreciadas. Este estudo avaliou o crescimento micelial vertical, a eficiência biológica, a produção e a composição nutricional de P. ostreatus produzido em substratos formulados com a combinação de resíduos do processamento de frutos de dendê (mesocarpo do fruto de dendê ­ MFD) e de amêndoas de cacau (tegumento de amêndoas de cacau ­ TAC). Os substratos foram formulados com as seguintes proporções de MFD e TAC (%/%): S1: 86,4/9,6; S2: 76,8/19,2; S3: 67,2/28,8; S4: 57,6/38,4 e S5: 48,0/48,0. Os substratos também receberam 3% de carvão e 1% de carbonato de cálcio. Os substratos S1, S2, S3 e S4 foram superiores quanto ao crescimento micelial vertical. S2 promoveu os melhores resultados para eficiência biológica (148,8%) e produção (560.5 g·kg-1). Os cogumelos produzidos em todos os substratos apresentaram valores nutricionais promissores. Entretanto, os cogumelos produzidos com o substrato S2 apresentaram o maior conteúdo de proteína bruta. De modo geral, S1 é o substrato recomendado por resultar na maior produção de cogumelos ricos em nutrientes. A produção de P. ostreatus em substratos compostos por MFD e TAC representa uma boa alternativa para a reciclagem desses resíduos com potenciais benefícios econômicos e ecológicos para as regiões produtoras de dendê e cacau.(AU)

Effective diffusion coefficients and bioconversion rates of inhibitory compounds in flocs of Saccharomyces cerevisiae

Background: Fermentation strategies for bioethanol production that use flocculating Saccharomyces cerevisiae yeast need to account for the mechanism by which inhibitory compounds, generated in the hydrolysis of lignocellulosic materials, are tolerated and detoxified by a yeast floc. Results: Diffusion coefficients and first-order kinetic bioconversion rate coefficients were measured for three fermentation inhibitory compounds (furfural, hydroxymethylfurfural, and vanillin) in self-aggregated flocs of S. cerevisiae NRRL Y-265. Thièle-type moduli and internal effectiveness factors were obtained by simulating a simple steady-state spherical floc model. Conclusions: The obtained values for the Thiéle moduli and internal effectiveness factors showed that the bioconversion rate of the inhibitory compounds is the dominant phenomenon over mass transfer inside the flocs.

Research progress on clinical gene polymorphism of clopidogrel resistance / 中国药学杂志

Clopidogrel plays an important role in anti-platelet aggregation, especially in acute coronary syndrome and percutaneous coronary intervention. However, clopidogrel resistance is common in clinical treatment. There are many factors response to clopidogrel resistance.Current researches concentrated in CYP450 enzyme gene polymorphism with clopidogrel resistance. There are few reviews on genetic polymorphisms of transporter and receptor binding sites, furthermore, the gene polymorphisms among different ethnic groups in plateau populations are very rare. In this paper, we mainly reviewed the relationship between gene polymorphism of drug-transporter and bioavailability of clopidogrel, the whole process of drug-metabolizing enzyme′s bioconversion of clopidogrel and the active metabolite of clopidogrel combine with the receptors.A mutation in the ABCB1 gene of the transporter was found to affect the bioavailability of clopidogrel. The key role of polymorphisms in the metabolic enzyme gene is CYP2C19 and CES1. The dose should be adjusted according to genotyping. The biologically active gene P2Y12 polymorphism affects the efficacy of clopidogrel. Therefore, understanding the clopidogrel gene polymorphism influencing factors can help individualized administration of clopidogrel to minimize thrombotic events caused by insufficient antiplatelet effect or hemorrhagic events caused by excessive anti-platelet effect.

Bottlenecks and modification strategies of 1,3-propanediol biosynthesis from glycerol / 生物工程学报

Crude glycerol is the main by-product of biodiesel production. A few microorganisms can transfer crude glycerol to 1,3-propanediol (1,3-PD) that is an important chemical material. There exist many limitations such as substrate inhibition, product inhibition when wild strains are used in 1,3-PD biosynthesis. In this review, based on the microbial transformation of 1,3-propanediol from glycerol and its limitations, some strategies using genetic engineering such as knockout or gene overexpression were summarized. The latest research progresses in biosynthesis of 1,3-propanediol from glycerol by genetically engineered strains are discussed.

Individualized clinical treatment from the prospective of hepatotoxicity of non-toxic traditional Chinese medicine / 中国中药杂志

Traditional Chinese medicine has a long history in clinical application, and been proved to be safe and effective. In recent years, the toxicity and side-effects caused by the western medicine have been attracted much attention. As a result, increasing people have shifted their attention to traditional Chinese medicine. Nonetheless, due to the natural origin of traditional Chinese medicine and the lack of basic knowledge about them, many people mistakenly consider the absolute safety of traditional Chinese medicine, except for well-known toxic ones, such as arsenic. However, according to the clinical practices and recent studies, great importance shall be attached to the toxicity of non-toxic traditional Chinese medicine, in particular the hepatotoxicity. Relevant studies indicated that the toxicity of non-toxic traditional Chinese medicine is closely correlated with individual gene polymorphism and constitution. By discussing the causes and mechanisms of the hepatotoxicity induced by non-toxic traditional Chinese medicine in clinical practices, we wrote this article with the aim to provide new ideas for individualized clinical therapy of traditional Chinese medicine and give guidance for rational and safe use of traditional Chinese medicine.

Over-expression of Mycobacterium neoaurum 3-ketosteroid-∆ 1-dehydrogenase in Corynebacterium crenatum for efficient bioconversion of 4-androstene-3, 17-dione to androst-1, 4-diene-3, 17-dione

Background: 3-Ketosteroid-∆¹-dehydrogenase (KSDD), a flavoprotein enzyme, catalyzes the bioconversion of 4-androstene-3,17-dione (AD) to androst-1,4-diene-3,17-dione (ADD). To date, there has been no report about characterization of KSDD from Mycobacterium neoaurum strains, which were usually employed to produce AD or ADD by fermentation. Results: In this work, Corynebacterium crenatum was chosen asa new host for heterologous expression of KSDD from M. neoaurum JC-12 after codon optimization of the KSDD gene. SDS-PAGE and western blotting results indicated that the recombinant C. crenatum harboring the optimized ksdd (ksdd n) gene showed significantly improved ability to express KSDD. The expression level of KSDD was about 1.6-fold increased C. crenatum after codon optimization. After purification of the protein, we first characterized KSDD from M. neoaurum JC-12, and the results showed that the optimum temperature and pH for KSDD activity were 30°C and pH 7.0, respectively. The Km and Vmax values of purified KSDD were 8.91 µM and 6.43 mM/min. In this work, C. crenatum as a novel whole-cell catalyst was also employed and validated for bioconversion of AD to ADD. The highest transformation rate of AD to ADD by recombinant C. crenatum was about 83.87% after 10 h reaction time, which was more efficient than M. neoaurum JC-12 (only 3.56% at 10 h). Conclusions: In this work, basing on the codon optimization, overexpression, purification and characterization of KSDD, we constructed a novel system, the recombinant C. crenatum SYPA 5-5 expressing KSDD, to accumulate ADDfromADefficiently. This work provided new insights into strengthening sterol catabolism by overexpressing the key enzyme KSDD, for efficient ADD production.

Consolidated briefing of biochemical ethanol production from lignocellulosic biomass

Bioethanol production is one pathway for crude oil reduction and environmental compliance. Bioethanol can be used as fuel with significant characteristics like high octane number, low cetane number and high heat of vaporization. Its main drawbacks are the corrosiveness, low flame luminosity, lower vapor pressure, miscibility with water, and toxicity to ecosystems. One crucial problem with bioethanol fuel is the availability of raw materials. The supply of feedstocks for bioethanol production can vary season to season and depends on geographic locations. Lignocellulosic biomass, such as forest-based woody materials, agricultural residues and municipal waste, is prominent feedstock for bioethanol cause of its high availability and low cost, even though the commercial production has still not been established. In addition, the supply and the attentive use of microbes render the bioethanol production process highly peculiar. Many conversion technologies and techniques for biomass-based ethanol production are under development and expected to be demonstrated. In this work a technological analysis of the biochemical method that can be used to produce bioethanol is carried out and a review of current trends and issues is conducted.

Bioconverted Jeju Hallabong tangor (Citrus kiyomi × ponkan) peel extracts by cytolase enhance antioxidant and anti-inflammatory capacity in RAW 264.7 cells

BACKGROUND/OBJECTIVES: Citrus and its peels have been used in Asian folk medicine due to abundant flavonoids and usage of citrus peels, which are byproducts from juice and/or jam processing, may be a good strategy. Therefore, the aim of this study was to examine antioxidant and anti-inflammatory effects of bioconversion of Jeju Hallabong tangor (Citrus kiyomi × ponkan; CKP) peels with cytolase (CKP-C) in RAW 264.7 cells. MATERIALS/METHODS: Glycosides of CKP were converted into aglycosides with cytolase treatment. RAW 264.7 cells were pre-treated with 0, 100, or 200 µg/ml of citrus peel extracts for 4 h, followed by stimulation with 1 µg/ml lipopolysaccharide (LPS) for 8 h. Cell viability, DPPH radical scavenging activity, nitric oxide (NO), and prostagladin E2 (PGE2) production were examined. Real time-PCR and western immunoblotting assay were performed for detection of mRNA and/or protein expression of pro-inflammatory mediators and cytokines, respectively. RESULTS: HPLC analysis showed that treatment of CKP with cytolase resulted in decreased flavanone rutinoside forms (narirutin and hesperidin) and increased flavanone aglycoside forms (naringenin and hesperetin). DPPH scavenging activities were observed in a dose-dependent manner for all of the citrus peel extracts and CKP-C was more potent than intact CKP. All of the citrus peel extracts decreased NO production by inducible nitric oxide synthase (iNOS) activity and PGE2 production by COX-2. Higher dose of CKP and all CKP-C groups significantly decreased mRNA and protein expression of LPS-stimulated iNOS. Only 200 µg/ml of CKP-C markedly decreased mRNA and protein expression of cyclooxygenase-2 in LPS-stimulated RAW 264.7 cells. Both 100 and 200 µg/ml of CKP-C notably inhibited mRNA levels of interleukin-1β (IL-1β) and IL-6, whereas 200 µg/ml CKP-C significantly inhibited mRNA levels of TNF-α. CONCLUSIONS: This result suggests that bioconversion of citrus peels with cytolase may enrich aglycoside flavanones of citrus peels and provide more potent functional food materials for prevention of chronic diseases attributable to oxidation and inflammation by increasing radical scavenging activity and suppressing pro-inflammatory mediators and cytokines.

Microspheres immobilized snailase prepared by method of microemulsion-coacervation for transform icariin / 中草药

Objective: To prepare microspheres immobilized snailase, then optimize the method for making microspheres immobilized snailase and the process conditions for transformation of icariin. Methods: The snailase was embedded in barium-alginate by the method of microemulsion-coacervation. The preparation process was optimized by orthogonal test. Moreover, to optimize and compare the conditions for transforming icariin, such as temperature, pH values, ratio of substrate, and concentration, and to investigate the properties of snailase. Results: Sodium alginates (1.0%), BaCl2 (1.0%), and emulgator were used for making microspheres immobilized snailase by emulsified and immobilized method. Compared with the free snailase, the stability of temperature and pH value of microspheres immobilized snailase was improved. The optimized conditions of microspheres immobilized snailase were 50℃, pH 5, microspheres immobilized snailase mass ratio of 3:1, icariin concentration of 0.4 mg/mL, conversion 2 h, conversion 3 times, transformational ratio up to (53.84±3.41)%. Conclusion: Microspheres immobilized snailase is stable and reusable. Besides, the process of transform icariin is moderate, so it is important and fundamental for industrialization.

Bioconversion of Citrus unshiu peel extracts with cytolase suppresses adipogenic activity in 3T3-L1 cells

BACKGROUND/OBJECTIVES: Citrus flavonoids have a variety of physiological properties such as anti-oxidant, anti-inflammation, anti-cancer, and anti-obesity. We investigated whether bioconversion of Citrus unshiu with cytolase (CU-C) ameliorates the anti-adipogenic effects by modulation of adipocyte differentiation and lipid metabolism in 3T3-L1 cells. MATERIALS/METHODS: Glycoside forms of Citrus unshiu (CU) were converted into aglycoside forms with cytolase treatment. Cell viability of CU and CU-C was measured at various concentrations in 3T3L-1 cells. The anti-adipogenic and lipolytic effects were examined using Oil red O staining and free glycerol assay, respectively. We performed real time-polymerase chain reaction and western immunoblotting assay to detect mRNA and protein expression of adipogenic transcription factors, respectively. RESULTS: Treatment with cytolase decreased flavanone rutinoside forms (narirutin and hesperidin) and instead, increased flavanone aglycoside forms (naringenin and hesperetin). During adipocyte differentiation, 3T3-L1 cells were treated with CU or CU-C at a dose of 0.5 mg/ml. Adipocyte differentiation was inhibited in CU-C group, but not in CU group. CU-C markedly suppressed the insulin-induced protein expression of CCAAT/enhancer-binding protein alpha (C/EBPalpha) and peroxisome proliferator-activated receptor gamma (PPARgamma) as well as the mRNA levels of CEBPalpha, PPARgamma, and sterol regulatory element binding protein 1c (SREBP1c). Both CU and CU-C groups significantly increased the adipolytic activity with the higher release of free glycerol than those of control group in differentiated 3T3-L1 adipocytes. CU-C is particularly superior in suppression of adipogenesis, whereas CU-C has similar effect to CU on stimulation of lipolysis. CONCLUSIONS: These results suggest that bioconversion of Citrus unshiu peel extracts with cytolase enhances aglycoside flavonoids and improves the anti-adipogenic metabolism via both inhibition of key adipogenic transcription factors and induction of adipolytic activity.

The Bioconversion of Red Ginseng Ethanol Extract into Compound K by Saccharomyces cerevisiae HJ-014

A beta-glucosidase producing yeast strain was isolated from Korean traditional rice wine. Based on the sequence of the YCL008c gene and analysis of the fatty acid composition, the isolate was identified as Saccharomyces cerevisiae strain HJ-014. S. cerevisiae HJ-014 produced ginsenoside Rd, F2, and compound K from the ethanol extract of red ginseng. The production was increased by shaking culture, where the bioconversion efficiency was increased 2-fold compared to standing culture. The production of ginsenoside F2 and compound K was time-dependent and thought to proceed by the transformation pathway of: red ginseng extract-->Rd-->F2-->compound K. The optimum incubation time and concentration of red ginseng extract for the production of compound K was 96 hr and 4.5% (w/v), respectively.

Bioconversion of Ginsenosides from Red Ginseng Extract Using Candida allociferrii JNO301 Isolated from Meju

Red ginseng (Panax ginseng), a Korean traditional medicinal plant, contains a variety of ginsenosides as major functional components. It is necessary to remove sugar moieties from the major ginsenosides, which have a lower absorption rate into the intestine, to obtain the aglycone form. To screen for microorganisms showing bioconversion activity for ginsenosides from red ginseng, 50 yeast strains were isolated from Korean traditional meju (a starter culture made with soybean and wheat flour for the fermentation of soybean paste). Twenty strains in which a black zone formed around the colony on esculin-yeast malt agar plates were screened first, and among them 5 strains having high beta-glucosidase activity on p-nitrophenyl-beta-D-glucopyranoside as a substrate were then selected. Strain JNO301 was finally chosen as a bioconverting strain in this study on the basis of its high bioconversion activity for red ginseng extract as determined by thin-layer chromatography (TLC) analysis. The selected bioconversion strain was identified as Candida allociferrii JNO301 based on the nucleotide sequence analysis of the 18S rRNA gene. The optimum temperature and pH for the cell growth were 20~30degrees C and pH 5~8, respectively. TLC analysis confirmed that C. allociferrii JNO301 converted ginsenoside Rb1 into Rd and then into F2, Rb2 into compound O, Rc into compound Mc1, and Rf into Rh1. Quantitative analysis using high-performance liquid chromatography showed that bioconversion of red ginseng extract resulted in an increase of 2.73, 3.32, 33.87, 16, and 5.48 fold in the concentration of Rd, F2, compound O, compound Mc1, and Rh1, respectively.