Dioscorea Zingiberensis C.H. Wright Research Articles

The Isolation, Identification and Immobilization Method of Three Novel Enzymes with Diosgenin-Producing Activity Derived from an Aspergillus flavus.

Diosgenin is an important raw material used in the synthesis of steroid drugs, and it is widely used in the pharmaceutical industry. The traditional method of producing diosgenin is through using raw materials provided via the plant Dioscorea zingiberensis C. H. Wright (DZW), which is subsequently industrially hydrolyzed using a high quantity of hydrochloric and sulfuric acids at temperatures ranging from 70 °C to 175 °C. This process results in a significant amount of unmanageable wastewater, creates issues of severe environmental pollution and consumes high quantities of energy. As an alternative, the enzymolysis of DZW to produce diosgenin is an environmentally and friendly method with wide-ranging prospects for its application. However, there are still only a few enzymes that are suitable for production on an industrial scale. In this study, three new key enzymes, E1, E2, and E3, with a high conversion stability of diosgenin, were isolated and identified using an enzyme-linked-substrate autography strategy. HPLC-MS/MS identification showed that E1, a 134.45 kDa protein with 1019 amino acids (AAs), is a zinc-dependent protein similar to the M16 family. E2, a 97.89 kDa protein with 910 AAs, is a type of endo-β-1,3-glucanase. E3, a 51.6 kDa protein with 476 AAs, is a type of Xaa-Pro aminopeptidase. In addition, the method to immobilize these proteins was optimized, and stability was achieved. The results show that the optimal immobilization parameters are 3.5% sodium alginate, 3.45% calcium chloride concentration, 1.4 h fixed time, and pH 8.8; and the recovery rate of enzyme activity can reach 43.98%. A level of 70.3% relative enzyme activity can be obtained after employing six cycles of the optimized technology. Compared with free enzymes, immobilized enzymes have improved stability, acid and alkaline resistance and reusability, which are conducive to large-scale industrial production.

Efficient alcoholysis of total saponins in D. zingiberensis C. H. Wright with novel magnetic solid acids through a cleaner approach

From the perspective of full utilization of biomass, a clean approach for the production of diosgenin from total saponins in Dioscorea zingiberensis C.H. Wright (DZW) was realized by preliminary separation and extraction. An alcoholysis approach was adopted to obtain diosgenin using a novel magnetic solid acid Fe3O4 @SiO2-Pr-S-Pr-SO3H Pr = -(CH2)3-) as the catalyst. The prepared solid acid has been fully characterized. The single factor methodology was used to optimize the process parameters. The optimum conditions were: 0.50 g Fe3O4 @SiO2-Pr-S-Pr-SO3H, 6.0 mL ethanol, the reaction temperature 120 ℃, reaction time 7.0 h and the maximum yield of diosgenin was 24.18% which are higher than conventional hydrolysis with 1.5 M sulfuric acid indicating the high catalytic activity of the solid acids. In addition, the solid acid had good reusability. The LC-MS technique was applied to analyze the main steroidal saponins in total saponins and possible mechanisms and pathways of hydrolysis and alcoholysis processes. Alcoholysis by an as-prepared magnetic solid acid is clearly excellent in high efficiency and reusability. It is a promising strategy for the green production of diosgenin through a cleaner approach.

Efficient degradation of solid residues from the diosgenin clean production industry to produce bioethanol

In this study, the degradation and bio-conversion of Dioscorea zingiberensis C. H. Wright solid residues (DZWSR) were performed. A large number of solid residues from Dioscorea zingiberensis C. H. Wright (DZW) are discarded or burned directly in the diosgenin industry, resulting in a serious waste of resources and environmental pollution. After dilute acid impregnation with low-pressure steam expansion (DALPSE), organic solvent extraction, enzyme hydrolysis, and other treatments, the hemicellulose degradation rate reached 94.71 %, the cellulose degradation rate increased to 88.29 %, and the fermentable sugar liquor was obtained. Compared with glucose fermentation, the ethanol yield by DZWSR hydrolysate fermentation was 83.93 %. It was found that DALPSE and the subsequent lignin extraction could break the lignocellulosic structure of DZWSR, which could effectively improve the accessibility of cellulose. This research will enhance the sustainability of the diosgenin industry and similar industries, and promote the resource utilization of biomass solid waste.

Preparation and characterization of oregano essential oil-loaded Dioscorea zingiberensis starch film with antioxidant and antibacterial activity and its application in chicken preservation

In this study, abundant starch was separated from the industrial crop Dioscorea zingiberensis C.H. Wright (DZW), and a novel bioactive packaging film loaded with oregano essential oil (OEO) was prepared and characterized. NaClO solution worked as a bleacher to prepare uniform starch powder from DZW tubers. OEO was selected from among three essential oils of Labiatae family plants for its strongest antibacterial activity. After the addition of OEO into the starch-based film, the UV–vis shielding property and antioxidant activity were enhanced. Meanwhile, the films still have a considerable performance in transparency, mechanical strength and water vapor permeability after incorporated with OEO. Furthermore, the 3% OEO-loaded starch film exhibited the strongest antibacterial activity against Bacillus subtilis, Escherichia coli and Staphylococcus aureus. It effectively lowered the total viable count of fresh chicken under 4 °C preservation conditions. These results revealed that the OEO-loaded DZW starch film can exert a positive effect on maintaining the quality and extending the shelf life of fresh meat. Therefore, readily accessible DZW tubers and oregano are very promising resources for application in degradable bioactive packaging film.

Ultrasound-assisted deep eutectic solvent/petroleum ether biphase system for the green and efficient production of diosgenin

Diosgenin, as an important precursor of steroid hormone drugs, is distributed in the root of Dioscorea Zingiberensis C. H. Wright (DZW) in the form of its sugar ligand-dioscin. The traditional production method of diosgenin is using sulfuric acid to hydrolyze the DZW, during the hydrolysis process, all components of the raw materials are degraded, resulting in a large amount of acid-containing wastewater with high Chemical Oxygen Demand (COD) values, causing serious pollution of the environment. In this paper, we proposed a biphase system composed of B acid type deep eutectic solvent (DES) and petroleum ether which can combine the extraction and hydrolysis of dioscin, and the purification of diosgenin into one step. The yield of diosgenin obtained by this method can reach 99.5% of that of the traditional method, while the COD value in the wastewater are less than 1% of the traditional method. The results showed that the best composition of DES is benzyltrimethylammonium chloride and oxalic acid, and the optimized parameters were material-to-liquid ratio 1:6 (w/v), ultrasonic power 200W, temperature 40 °C, ultrasonic time 15min and DES/petroleum ether ratio 1:6(w/v). After repeated use of the bi-phase system for 6 times, the production efficiency of diosgenin can still reach more than 95%, and the COD value of wastewater were stable. Furthermore, in order to better understand the advantages of this method, this method was compared with several other diosgenin production methods. In summary, the DES/petroleum ether biphase system proposed in this paper is an efficient and cleaner production method of diosgenin. • DES is simple to prepare and easy to recycle. • The DES/petroleum ether biphase system used in this study couples the production and purification of diosgenin into one step. • The yield and purity of diosgenin obtained in this process is comparable to that of the traditional method, while the COD value in the wastewater is less than 1% of the value of the traditional method.

Identification and a phased pH control strategy of diosgenin bio-synthesized by an endogenous Bacillus licheniformis Syt1 derived from Dioscorea zingiberensis C. H. Wright.

Diosgenin is widely used as one precursor of steroidal drugs in pharmaceutical industry. Currently, there is no choice but to traditionally extract diosgenin from Dioscorea zingiberensis C. H. Wright (DZW) or other plants. In this work, an environmentally friendly approach, in which diosgenin can be bio-synthesized by the endophytic bacterium Bacillus licheniformis Syt1 isolated from DZW, is proposed. Diosgenin produced by the strain was identified by high-performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR), and Fourier transform infrared spectroscopy (FTIR). The thermal gravimetric analysis (TGA) showed that the melting point of the diosgenin product was 204°C. The optical rotation measurement exhibited that the optical rotation was α20589 = - 126.1° ± 1.5° (chloroform, c = 1%): negative sign means that the product is left-handed, which is very important to further produce steroid hormone drugs. Cholesterol may be the intermediate product in the diosgenin biosynthesis pathway. In the batch fermentation process to produce diosgenin using the strain, pH values played an important role. A phased pH control strategy from 5.5 to 7.5 was proved to be more effective to improve production yield than any single pH control, which could get the highest diosgenin yield of 85 ± 8.6 mg L-1. The proposed method may replace phyto-chemistry extraction to produce diosgenin in the industry in the future.Key points• An endophytic Bacillus licheniformis Syt1 derived from host can produce diosgenin.• A dynamic pH industrial control strategy is better than any single pH control.• Proposed diosgenin-produced method hopefully replaces phyto-chemistry extraction.

Polymer‐based solid acid catalyst for the green production of diosgenin

AbstractIn this study, a novel polymer‐based solid acid catalyst was prepared by free radical polymerization of N, N′‐methylenebisacrylamide (MBA) and allyl sodium sulfonate (SAS), followed by acidifying of diluted hydrochloric acid. The prepared solid acid was fully characterized by FTIR, solid‐state 13C‐NMR, TGA, XPS, element analysis, and SEM. In addition, the prepared solid acid poly(MBA‐SAA) was used as a catalyst to extract diosgenin from Dioscorea zingiberensis C.H.Wright (DZW) under solvethermal condition using ethanol as a solvent. And the optimal conditions of temperature (104.02°C), alcoholysis time (5.87 h) and solid acid dosage (0.39 g), were obtained by response surface analysis based on the results of single factor experiments. The yield of diosgenin (14.53%) obtained in the alcoholysis is higher than that of the traditional sulfuric acid hydrolysis method. The alcoholysis products were analyzed by high performance liquid chromatography‐mass spectrometry (LC–MS) and UV–vis, respectively. The results showed that the solid acid with higher catalytic activity and good reusability can replace sulfuric acid to extract diosgenin, which has a great potential for the clean production of diosgenin in industry.

Screening and Selection of a New Medium for Diosgenin Production via Microbial Biocatalysis of Fusarium sp.

Steroidal saponins are widely used as starting precursors and medical intermediates for the semi-/total-synthesis of hundreds of steroidal drugs. One such steroidal saponin is diosgenin, which has attracted significant attention due to the huge market demand in the pharmaceutical industry. Due to water waste and severe environmental pollution, the traditional diosgenin production process based on direct acid hydrolysis is no longer used. In this study, to develop a submerged fermentation (SmF) medium for clean diosgenin production via efficient microbial biocatalysis, the Box–Behnken design (BBD) in combination with the Plackett–Burman design (PBD) was used to determine the medium compositions for Fusarium strains. Three components (wheat bran, phosphate, and Tween-80) were determined as significant factors by the PBD. Using the BBD, the three significant factors were further optimized, and the optimum values were determined for maximal diosgenin production. With 21.16 g/L of wheat bran, 9.60 g/L of phosphate, and 1.97 g/L of Tween-80, the diosgenin yield was 2.28%, i.e., 3.17 mg/L/h. The experimental values agreed with the predicted values, representing a significant increase in diosgenin production compared to its production using the basic SmF medium. For the first time, we reported the development of a new medium for Fusarium strains to produce diosgenin via microbial biocatalysis of the root of Dioscorea zingiberensis C. H. Wright (DZW). A simple-composition, low-cost, and high-efficiency medium was developed for the first time for the SmF of Fusarium strains. The medium is considered useful for large-scale SmF and may be applicable to other fungi. This study lays a solid foundation for diosgenin production in an acid-free and wastewater-free way. It may also provide fundamental support for producing other value-added products via microbial biocatalysis of low-value materials by endophytic fungi.

Development of a New Bioprocess for Clean Diosgenin Production through Submerged Fermentation of an Endophytic Fungus.

Diosgenin is used widely to synthesize steroidal hormone drugs in the pharmaceutical industry. The conventional diosgenin production process, direct acid hydrolysis of the root of Dioscorea zingiberensis C. H. Wright (DZW), causes large amounts of wastewater and severe environmental pollution. To develop a clean and effective method, the endophytic fungus Fusarium sp. CPCC 400226 was screened for the first time for the microbial biotransformation of DZW in submerged fermentation (SmF). Statistical design and response surface methodology (RSM) were implemented to develop the diosgenin production process using the Fusarium strains. The environmental variables that significantly affected diosgenin yield were determined by the two-level Plackett–Burman design (PBD) with nine factors. PBD indicates that the fermentation period, culture temperature, and antifoam reagent addition are the most influential variables. These three variables were further optimized using the response surface design (RSD). A quadratic model was then built by the central composite design (CCD) to study the impact of interaction and quadratic effect on diosgenin yield. The values of the coefficient of determination for the PBD and CCD models were all over 0.95. P-values for both models were 0.0024 and <0.001, with F-values of ∼414 and ∼2215, respectively. The predicted results showed that a maximum diosgenin yield of 2.22% could be obtained with a fermentation period of 11.89 days, a culture temperature of 30.17 °C, and an antifoam reagent addition of 0.20%. The experimental value was 2.24%, which was in great agreement with predicted value. As a result, over 80% of the steroidal saponins in DZW were converted into diosgenin, presenting a ∼3-fold increase in diosgenin yield. For the first time, we report the SmF of a Fusarium strain used to produce diosgenin through the microbial biotransformation of DZW. A practical diosgenin production process was established for the first time for Fusarium strains. This bioprocess is acid-free and wastewater-free, providing a promising environmentally friendly alternative to diosgenin production in industrial applications. The information provided in the current study may be applicable to produce diosgenin in SmF by other endophytic fungi and lays a solid foundation for endophytic fungi to produce natural products.

Green Conversion of Saponins to Diosgenin in an Alcoholysis System Catalyzed by Solid Acid Derived from Phosphorus Tailings.

This work proposed to prepare solid acid from phosphorus tailings and successfully convert Dioscorea zingiberensis C.H. Wright (DZW) into diosgenin from the perspective of solid waste resource reuse and clean production. The results showed that SiO2–SO3H solid acid could catalyze the production of diosgenin from total saponins under solvothermal reaction conditions. In addition, the parameters of a single factor, such as the amount of SiO2–SO3H, solvent volume, reaction temperature, and reaction time, were optimized to confirm the optimal range of reaction conditions, and the optimal process conditions were determined by the response surface method. The yield of diosgenin was 2.45 ± 0.17% under the optimum conditions, and the yield of diosgenin was increased by 12.90% compared with the traditional acid hydrolysis process. Except the relatively higher catalytic activity, the alcoholysis approach for the production of diosgenin has no waste liquid to discharge. The products were analyzed by high-performance liquid chromatography-mass spectrometry, and the pathway to convert total saponins into diosgenin under SiO2–SO3H has been proposed. Moreover, the adopted catalyst can be prepared with very low cost from phosphorus tailings. Considering the obvious superiorities, the alcoholysis approach in this work could be a promising strategy for green production of diosgenin as well as a possible utilization pathway of phosphorus tailings.

Preparation of poly (styrene‐co‐allyl sulfonic acid) as a novel catalyst for the alcoholysis of dioscin

This work describes the preparation of a novel solid acid, poly (styrene‐co‐allyl sulfonic acid) (St‐SAA), and its activity test for extracting diosgenin from Dioscorea Zingiberensis C.H. Wright (DZW). The solid acid was synthesized by the radical polymerization of styrene and sodium allyl sulfonate in water/ethanol solution, followed by the acidification. The prepared solid acid was characterized by infrared spectrum (FTIR), thermogravimetric analysis (TGA), X‐ray photoelectron spectroscopy (XPS), element analysis, and scanning electron microscope. The solid acid catalyst was used to extract diosgenin from DZW. It was found that the alcoholysis activity of the solid acid was higher than the hydrolysis activity of sulfuric acid with the same mass under the same conditions. Besides, the solid acid can be separated from the reaction mixture for the recycling extraction of diosgenin, and the solid acid has no loss of mass and no decrease of alcoholysis activity after repeated cycles.

Eco-friendly microbial production of diosgenin from saponins in Dioscorea zingiberensis tubers in the presence of Aspergillus awamori

A novel microbial procedure was proposed for diosgenin production from Dioscorea zingiberensis C.H. Wright (DZW) tubers via employing Aspergillus awamori for the first time. The optimal conditions of fermenter cultivation were established as inoculation dosage of 8%, fermentation temperature of 30 °C, cultivation time of 8 days, initial pH of 7.0 and a stirring rate of 180 rpm when the converted diosgenin content reached a peak value of 74.26 ± 3.23 mg/g substrate. The product was purified by silica gel column and then confirmed as diosgenin (purity: 96.9 ± 2.42%) by nuclear magnetic resonance (NMR). Compared with traditional acid hydrolysis, this new process generated indeed less wastewater with lower chemical oxygen demand (COD) reduced to 500 mg/L from 10,000 mg/L and absence of acid and alkali. This research provided definitely an environmental and high-efficiency microbial technology for diosgenin production.

Isolation of endophytic fungi from Dioscorea zingiberensis C. H. Wright and application for diosgenin production by solid-state fermentation.

In this study, endophytic fungi were isolated from Dioscorea zingiberensis C.H. Wright (DZW), and a novel clean process to prepare diosgenin from DZW was developed. A total of 123 strains of endophytic fungi were isolated from different plant tissues of DZW. Among them, the strain Fusarium sp. (CPCC 400709) showed the best activity of hydrolyzing steroidal saponins in DZW into diosgenin. Thus, this strain was used to prepare diosgenin from DZW by solid-state fermentation. The fermentation parameters were optimized using response surface methodology, and a high yield of diosgenin (2.16%) was obtained at 14.5% ammonium sulfate, an inoculum size of 12.3%, and 22days of fermentation. Furthermore, the highest diosgenin yield (2.79%) was obtained by co-fermentation with Fusarium sp. (CPCC 400709) and Curvularia lunata (CPCC 400737), which was 98.9% of that obtained by β-glucosidase pretreated acid hydrolysis (2.82%). This process is acid-free and wastewater-free, and shows promise as an effective and clean way to prepare diosgenin for use in industrial applications from DZW.

A novel glycosylated solution from Dioscorea zingiberensis C.H. Wright significantly improves the solvent productivity of Clostridium beijerinckii

The economics of bio-solvent production are largely dependent on the cost of the fermentation substrate. Dioscorea zingiberensis C.H. Wright (DZW), the main raw material used to produce saponin, contains 13–18% starch which can be directly saccharified to a saccharification liquid of DZW starch (SLDS) that contains abundant nutrients. In this study, the water-soluble micromolecule compounds in SLDS were quantified through 1H NMR. Using SLDS as the substrate to conduct ABE fermentation by Clostridium beijerinckii, the fermentation cycle was shortened 24h, the maximum biomass and consumption rate of the glucose significantly increased, and the productivity of total solvents were increased by 0.244±0.010g/L/h compared to standard P2 medium. Expression analysis of genes encoding key enzymes involved in acetone and butanol production and glucose consumption showed that they were induced by SLDS. Taken together, SLDS is a useful and renewable glycosylated solution for ABE fermentation.

Microwave-assisted extraction and a new determination method for total steroid saponins from Dioscorea zingiberensis C.H. Wright

An efficient microwave-assisted extraction (MAE) technique was applied to isolate total steroid saponins from Dioscorea zingiberensis C.H. Wright (DZW). The optimal extracting conditions were established as 75% ethanol as solvent, ratio of solid/liquid 1:20 (g/ml), temperature 75°C, irradiation power 600W and three extraction cycles of 6min each. Scanning electron microscopy (SEM) images of DZW processed by four different extractions provided visual evidence of the disruption effect on DZW. Diosgenin was quantified by HPLC and examined further by LC–ESI/MS after acid hydrolysis. Total steroid saponins were calculated using diosgenin from total steroid saponins. The MAE procedure was optimized, validated and compared with other conventional extraction processes. This report provides a convenient technology for the extraction and quantification of total saponins of DZW combining MAE with HPLC and LC–ESI/MS for the first time.

Conversion of furostanol saponins into spirostanol saponins improves the yield of diosgenin from Dioscorea zingiberensis by acid hydrolysis

Conversion of furostanol saponins into spirostanol saponins avoiding the side product increases the diosgenin yield in acid hydrolysis of Dioscorea zingiberensis.

Therapeutic effects of total steroid saponin extracts from the rhizome of Dioscorea zingiberensis C.H.Wright in Freund's complete adjuvant induced arthritis in rats.

The aim of our present study is to explore the anti-arthritic potential effect of total steroid saponins (TSSNs) extracted from the rhizome of Dioscorea zingiberensis C.H.Wright (DZW) and to investigate the underlying mechanisms. This work was performed using adjuvant-induced arthritis (AIA) rats in vivo and lipopolysaccharide (LPS) simulated 264.7 macrophage cells in vitro. In AIA-induced arthritic rats, TSSN significantly alleviated the arthritic progression through evaluating arthritic score, immune organ indexes, paw swelling, and body weight. This phenomenon was well correlated with significant suppression of the overproduction of inflammation cytokines (IL-1, IL-1β, IL-6, and TNF-α), oxidant stress makers (MDA and NO), eicosanoids (LTB4 and PGE2), and inflammatory enzymes (5-LOX and COX-2) versus the AIA rats without treatment. On the contrary, the release of SOD and IL-10 was profoundly increased. What's more, TSSN could obviously ameliorate the translocation of NF-κB to the nucleus through phosphorylation of the p65 and IκBα in vivo and in vitro. The current findings demonstrated that TSSN could protect the injured ankle joint from further deterioration and exert its satisfactory anti-arthritis properties through anti-inflammatory and anti-oxidant effects via inactivating the NF-κB signal pathway. This research implies that DZW may be a useful therapeutic agent for the treatment of human arthritis.

Quality control and identification of steroid saponins from Dioscorea zingiberensis C. H. Wright by fingerprint with HPLC-ELSD and HPLC-ESI-Quadrupole/Time-of-fight tandem mass spectrometry

In this study, a fingerprint of steroid saponins, the major bioactive constituents in the crude extracts from Dioscorea zingiberensis C. H. Wright (DZW), has been established for the first time by combined use of the following two methods: high-performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD) and the simultaneous characterization of the steroid saponins by high-performance liquid chromatography coupled with electrospray ionization-mass spectrometry and quadrupole tandem time-of-fight mass analyzers detection (HPLC-ESI-Q/TOF). All HPLC analyses were carried out on a Welchrom C18 column (250mm×4.6mm I.D., 5μm) with a mobile phase composed of water and acetonitrile under gradient elution. There were 68 common characteristic peaks in the fingerprints, in which 12 of them were confirmed by comparing their mass spectra and retention times with those of the reference compounds. In order to identify other unknown peaks, their fragmentation behaviors characteristic of the major groups of steroid saponins from DZW with six types of aglycone skeletons were discussed in detail, and possible MS/MS fragmentation pathways were proposed for aiding the structural identification of these components. According to the summarized fragmentation patterns, these peaks were tentatively assigned by matching their empirical molecular formula with those of the published compounds, or by elucidating their quasi-molecular ions and fragment ions referring to available literature information when the reference standards were unavailable. As a result, 22 new steroid saponins were found in DZW for the first time. In addition, the quantitative analysis of the nine (except for the reference compounds A, B, and C) known peaks was accomplished at the same time which indicated that there was a great variability in the amount of these active compounds in different batches in the crude extracts. This approach could demonstrate that the fingerprint could be considered to be a suitable tool to comprehensively improve the quality control of DZW. The identification and structural elucidation of the peaks in the fingerprint may provide important experimental data for further pharmacological and clinical researches.

Pathways of biotransformation of zingiberen newsaponin from Dioscorea zingiberensis C. H. Wright to diosgenin

Abstract A new steroidal saponin-β-glucosidase from Aspergillus flavus that specifically hydrolyzed the terminal β- d -glucosyl group at the C-3 position of zingiberen newsaponin, deltonin and trillin from Dioscorea zingiberensis C. H. Wright (DZW) was purified, and characterized. The optimal temperature and pH for the new steroidal saponin-β-glucosidase was 50 °C and pH 5.0. The steroidal saponin-β-glucosidase was stable at 30–60 °C, and retained more than 80% activity. Further, the purified protein was analyzed by ESI-Q-TOF proteomic analyzer. The results indicated that this enzyme is a β-glucosidase of the type glycosidase hydrolase 3 (GH3). Using a combination of the steroidal saponin-β-glucosidase and steroidal saponin-α-1,2-rhamnosidase from Curvularia lunata obtained previously in our lab, the saponins zingieren newsaponin and deltonin could be converted to diosgenin. The pathways of converting zingiberen newsaponin and deltonin into diosgenin by the two key enzymes were elucidated in this study.

Novel method utilizing microbial treatment for cleaner production of diosgenin from Dioscorea zingiberensis C.H. Wright (DZW)

A novel method utilizing microbial treatment for cleaner production of diosgenin from Dioscorea zingiberensis C.H. Wright (DZW) was presented. A new Bacillus pumilus HR19, which has the great ability to secrete pectinase, was screened and applied in the microbial treatment. Low-pressure steam expansion pretreatment (LSEP) was employed in advance to assist microbial treatment efficiently in releasing saponins, which are the precursors of diosgenin. Compared with the traditional process of acid hydrolysis, this novel process reduced the consumptions of water, acid and organic solvent by more than 92.5%, 97.0%, 97.0%, respectively, while simultaneously increasing the diosgenin yield by 6.21%. In addition, the microbial treatment was more efficient than enzymatic treatment, which arised from that microorganisms could be induced to secrete related enzymes by the compositions of DZW and relieve product inhibition by utilizing enzyme hydrolysates.