Varieties systematization and standards status analysis of fermented Chinese medicine / 中国中药杂志

Fermented Chinese medicine has long been used. Amid the advance for preservation of experience, the connotation of fermented Chinese medicine has been enriched and improved. However, fermented Chinese medicine prescriptions generally contain a lot of medicinals. The fermentation process is complicated and the conventional fermentation conditions fail to be strictly controlled. In addition, the judgment of the fermentation end point is highly subjective. As a result, quality of fermented Chinese medicine is of great difference among regions and unstable. At the moment, the quality standards of fermented Chinese medicine are generally outdated and different among regions, with simple quality control methods and lacking objective safe fermentation-specific evaluation indictors. It is difficult to comprehensively evaluate and control the quality of fermented medicine. These problems have aroused concern in the industry and also affected the clinical application of fermented Chinese medicine. This article summarized and analyzed the application, quality standards, and the modernization of fermentation technology and quality control methods of fermented Chinese medicine and proposed suggestions for improving the quality standards of the medicine, with a view to improving the overall quality of it.

Genetic modification of acetogens and optimization of fermentation process in C1-gas bioconversion / 生物工程学报

The current linear economy model relies on fossil energy and increases CO2 emissions, which contributes to global warming and environmental pollution. Therefore, there is an urgent need to develop and deploy technologies for carbon capture and utilization to establish a circular economy. The use of acetogens for C1-gas (CO and CO2) conversion is a promising technology due to high metabolic flexibility, product selectivity, and diversity of the products including chemicals and fuels. This review focuses on the physiological and metabolic mechanisms, genetic and metabolic engineering modifications, fermentation process optimization, and carbon atom economy in the process of C1-gas conversion by acetogens, with the aim to facilitate the industrial scale-up and carbon negative production through acetogen gas fermentation.

Isolation and Identification of Arisaema Cum Bile Fermentation Strain and Preliminary Investigation on Its Compound Strain Fermentation / 中国实验方剂学杂志

ObjectiveIn order to solve the problem that the quality and stability of Arisaema Cum Bile in the fermentation process with hybrid bacteria were not easy to control， the microorganism in the fermentation process of Arisaema Cum Bile was isolated and identified， the dominant strains were screened and the fermentation process of Arisaema Cum Bile with compound bacteria was investigated. MethodThe submerged culture during the fermentation process of Arisaema Cum Bile was taken out for strain separation and purification. Bacteria and fungi multiphase identification and detection methods and automatic microbial analysis system were used to analyze and compare DNA sequences and identify microorganisms. The isolated and identified strains were respectively inoculated and fermented. After screening the dominant strains， a preliminary exploration of compound strain fermentation were carried out. The contents of index components in Arisaema Cum Bile fermented by compound strain and traditional Arisaema Cum Bile were compared by ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry （UPLC-QqQ-MS/MS）. Mmobile phase was 0.1% formic acid acetonitrile solution （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-2 min， 35%-45%A； 2-10 min， 45%-48%A； 10-12 min， 48%-100%A； 12-12.01 min， 100%-35%A； 12.01-15 min， 35%-65%A）， the flow rate was set at 0.35 mL·min-1. The mass spectrographic analysis employed electrospray ionization （ESI）， negative ion acquisition mode and multiple reaction monitoring （MRM） scanning mode were adopted to collect information， the collection range was m/z 50-1 000. ResultEight microorganisms were isolated and identified from the submerged culture of Arisaema Cum Bile. Among them， Enterococcus sp. （anaerobic） and E. casseliflavus were selected as the dominant strains in the fermentation process. Compared with the traditional fermentation method， the contents of chenodeoxycholic acid， hyodeoxycholic acid and hyocholic acid in free cholic acid increased by 1.76， 0.06， 0.19 mg·g-1， respectively. In bound cholic acid， glycochenodeoxycholic acid， taurochenodeoxycholic acid， glycohyodeoxycholic acid， taurohyodeoxycholic acid， glycohyocholic acid， taurine porcine cholic acid decreased by 0.63， 0.23， 0.26， 0.16， 0.03， 0.04 mg·g-1， respectively. ConclusionArisaema Cum Bile with compound strain fermentation （Enterococcus sp. and E. casseliflavus） can be fermented more completely， the fermentation cycle can be shortened， and the quality and stability of products can be improved.

An Overview of Fermentation Technology Evolution and Process Control of Sojae Semen Praeparatum / 中国实验方剂学杂志

The historical evolution， fermentation technology and key links of Sojae Semen Praeparatum （SSP） were sorted out by consulting ancient books and modern literature， and the influencing factors and control methods of quality were analyzed and summarized in order to provide reference for the quality control of SSP. After analysis， it was found that in the fermentation process of SSP， fermentation strains， miscellaneous bacteria， temperature and humidity were all important factors affecting the quality of SSP. The condition control of "post fermentation" process has been paid more attention to in the past dynasties. In addition， the delicious SSP recognized in ancient times should be made from mold fermentation， and the breeding and application of fermented mold may be the key point to solve the quality problem of SSP. Therefore， based on the evaluation indexes of SSP in the past dynasties， it is of great significance to study and optimize the technological conditions such as strain， temperature and humidity in depth to improve the quality of SSP.

Distribution of archaeal community in the mud pit during strong-flavor baijiu fermentation / 生物工程学报

Multi-species solid-state fermentation in a mud pit is one of the typical features of strong-flavor baijiu, in which archaea plays important roles, however, the archaeal community distribution and diversity during fermentation are still lack of research. The biomass, composition and succession of archaea communities in fermented grains and pit mud were analyzed by high throughput sequencing. The potential interaction between archaea and bacteria was analyzed by co-occurrence network. Results demonstrate that the average biomass of archaea in pit mud was about 200 times higher than that of fermented grains. There was no significant difference in archaeal community structure between fermented grains and pit mud (r=0.017, P=0.074), but succession patterns between them showed significant correlation (r=0.30, P=0.03). Methanobacterium was the most abundant archaea in fermented grains and pit mud, and other dominant groups included Methanosarcina, Methanocorpusculum, Methanoculleus, and Methanobrevibacter. The co-occurrence network analysis showed that Methanobacterium was positively correlated with most bacteria in fermented grains and pit mud, especially with Hydrogenispora and Caproiciproducens, the dominant bacteria in pit mud. Our results revealed the temporal and spatial distribution characteristics and potential functions of the archaeal community in the mud pit of strong-flavor baijiu.

On-line monitoring of multiple component parameters during ethanol fermentation by near-infrared spectroscopy / 生物工程学报

The quantity of biomass, glucose concentration and ethanol concentration are important parameters in ethanol fermentation. Traditional methods are usually based on samples for off-line measurement, which not only requires multiple instruments for test and analysis but also consumes notable time and effort, and therefore is inconvenient for real-time process control and optimization. In this study, an in-situ detection method based on the near-infrared (NIR) spectroscopy is proposed for measuring the above process parameters in real time. The in-situ measurement is carried out by using an immersion type NIR spectroscopy. A multi-output prediction model for simultaneously estimating the quantity of glucose, biomass and ethanol is established based on a multi-output least-squares support vector regression algorithm. The experimental results show that the proposed method can precisely measure the quantity of glucose, biomass and ethanol during the ethanol fermentation process. Compared to the existing partial-least-squares method for modeling and prediction of individual components, the proposed method could evidently improve the measurement accuracy and reliability.

Development and application of in-situ microscopy in on-line monitoring of cell biomass / 生物工程学报

With the rapid development of modern biotechnology, fermentation process is increasingly important in industrial production. To guarantee the stability of products, fermentation process should be elaborately monitored and controlled. Biomass is an important parameter for on-line monitoring in bioprocesses because biomass can reflect cell growth in a bioreactor directly. In-situ microscope, a non-invasive and image-analysis based technology, can real-time monitor cells in biological process. This review summarizes the development and application of in-situ microscopy in biomass monitoring.

Preliminary analysis of primary and secondary factors of γ-aminobutyric acid enrichment in fermentation process of Sojae Semen Praeparatum / 中草药

Objective: To study the primary and secondary factors of γ-aminobutyric acid (GABA) enrichment in the processing of Sojae Semen Praeparatum (SSP), and lay the foundation for revealing the formation mechanism of high content of GABA in SSP. Methods: The dynamic changes of pH value, temperature, moisture, protease and glutamic acid decarboxylase (GAD) in the processing of SSP were determined by conventional methods. The GABA content of each sample was determined by pre-column on-line derivatization established by our laboratory. The correlation between each indicator and GABA was analyzed by SPSS software. Results: Correlation analysis and multiple regression analysis showed that the correlation coefficient between water, acid protease and GABA was 0.211 and -0.340, respectively, and the P values were 0.324 and 0.228, respectively. The correlation was small and there was no statistical difference. The absolute value of the regression coefficient showed that the primary and secondary status of other indicators in the formation of GABA was pH value (-0.375) > temperature (-0.284) > GAD (0.140) > alkaline protease (0.047) > neu-protease (-0.030), in which pH value, temperature and neutral protease had a negative correlation with GABA, and GAD activity and alkaline protease had a positive correlation with GABA. Conclusion: The temperature, pH value, GAD, neutral and alkaline protease are important factors affecting the formation of GABA in the fermentation process of SSP.

Scaling-up fermentation of Escherichia coli for production of recombinant P64k protein from Neisseria meningitidis

Background: P64k is a Neisseria meningitidis high molecular weight protein present in meningococcal vaccine preparations. The lpdA gene, which encodes for this protein, was cloned in Escherichia coli and the P64k recombinant protein was expressed in E. coli K12 GC366 cells under the control of a tryptophan promoter. P64k was expressed as an intracellular soluble protein about 28% of the total cellular protein. Several scale-up criteria of fermentation processes were studied to obtain the recombinant P64k protein at the pilot production scale. Results: The best operational conditions at a larger scale production of P64k recombinant protein were studied and compared using the four following criteria: Constant Reynold's number (Re constant), Constant impeller tip speed (n di constant), Constant power consumption per unit liquid volume (P/V constant) and Constant volumetric oxygen transfer coefficients (KLa/k constant). The highest production of the recombinant protein was achieved based on the constant KLa/k scale-up fermentation criterion, calculating the aeration rate (Q) and the impeller agitation speed (n) by iterative process, keeping constant the KLa/k value from bench scale. The P64k protein total production at the 50 l culture scale was 546 mg l -1 in comparison with the 284 mg l -1 obtained at 1.5 l bench scale. Conclusions: The methodology described herein, for the KLa/k scale-up fermentation criterion, allowed us to obtain the P64k protein at 50 l scale. A fermentation process for the production of P64k protein from N. meningitidis was established, a protein to be used in future vaccine formulations in humans.

Investigations on occupational hazards in a kitchen waste treatment plant / 中华劳动卫生职业病杂志

Objective@#To explore the distribution and intensity of kitchen waste treatment plant, providing scientific basis for occupational diseases prevention and treatment in that industry.@*Methods@#30 operating personnel in a kitchen waste disposal enterprise were selected as the research group and 16 non-operating personnel as the control group. The epidemiological survey was carried out in June 2015 and July 2016, and the results of occupational hazards and occupational health monitoring during 2015 to 2017 were analyzed.@*Results@#The results of various occupational hazards factors during 2015 to 2017 were all in accordance with the occupational health limit, but the results of occupational health examination were abnormal. The occupational correlation could not be ruled out by the abnormality of electrical audiometry, electrocardiogram and Department of Dermatology examination.@*Conclusion@#Workers in kitchen waste treatment plant are likely to suffer low-intensity, continuous health damage. There is occupational health risk in that industry. The accumulation of basic information and the improvement of relevant standards and methods are needed for further study.

On-line monitoring of biomass in 1,3-propanediol fermentation by Fourier-transformed near-infrared spectra analysis / 生物工程学报

Biomass is an important parameter reflecting the fermentation dynamics. Real-time monitoring of biomass can be used to control and optimize a fermentation process. To overcome the deficiencies of measurement delay and manual errors from offline measurement, we designed an experimental platform for online monitoring the biomass during a 1,3-propanediol fermentation process, based on using the fourier-transformed near-infrared (FT-NIR) spectra analysis. By pre-processing the real-time sampled spectra and analyzing the sensitive spectra bands, a partial least-squares algorithm was proposed to establish a dynamic prediction model for the biomass change during a 1,3-propanediol fermentation process. The fermentation processes with substrate glycerol concentrations of 60 g/L and 40 g/L were used as the external validation experiments. The root mean square error of prediction (RMSEP) obtained by analyzing experimental data was 0.341 6 and 0.274 3, respectively. These results showed that the established model gave good prediction and could be effectively used for on-line monitoring the biomass during a 1,3-propanediol fermentation process.

Biopharmaceuticals from microorganisms: from production to purification

ABSTRACT The use of biopharmaceuticals dates from the 19th century and within 5-10 years, up to 50% of all drugs in development will be biopharmaceuticals. In the 1980s, the biopharmaceutical industry experienced a significant growth in the production and approval of recombinant proteins such as interferons (IFN α, β, and γ) and growth hormones. The production of biopharmaceuticals, known as bioprocess, involves a wide range of techniques. In this review, we discuss the technology involved in the bioprocess and describe the available strategies and main advances in microbial fermentation and purification process to obtain biopharmaceuticals.

Biopharmaceuticals from microorganisms: from production to purification

ABSTRACT The use of biopharmaceuticals dates from the 19th century and within 5-10 years, up to 50% of all drugs in development will be biopharmaceuticals. In the 1980s, the biopharmaceutical industry experienced a significant growth in the production and approval of recombinant proteins such as interferons (IFN , , and ) and growth hormones. The production of biopharmaceuticals, known as bioprocess, involves a wide range of techniques. In this review, we discuss the technology involved in the bioprocess and describe the available strategies and main advances in microbial fermentation and purification process to obtain biopharmaceuticals.

Effect of secondary fermentation on processing technology for fermented soybeans / 中草药

Objective: To optimize the fermentation processing technology for fermented soybeans, and to identify that the secondary fermentation process is an important link in fermentation processing of fermented soybeans. Methods: According to Chinese Pharmacopoeia 2010, combined with the ancient way to produce fermented soybeans, the contents of total isoflavones, daidzein, and genistein were as chemical indexes, the finished product properties (color, smell, degree of shrinkage, cross section, and hardness) were as organoleptic quality indexes, and the processing parameters before and within the secondary fermentation process, including cooking time, fermentation temperature, fermentation time, secondary fermentation temperature and time were optimized. Results: The optimal processing technology was as follows: soybeans were boiled for 1.5 h after absorbing drug juice, and then fermented for 6-8 d at (30 ± 2)°C until yellow cladding. After washing off yellow cladding, soybeans were placed in the container, sealed using water, and put into secondary fermentation process. The container was in secondary fermentation for 12-15 d at (30 ± 2)°C. During the period of secondary fermentation, the soybeans were poured out every 3 d with stiring and slightly drying for four to five times, and at last was slightly steamed and dryed. The quality of fermented soybeans after secondary fermentation had more obvious advantage than that without secondary fermentation. The finished product had aromatic odor with light color and grain soft. The cross section color was brownish black and hand skin shrivel. The contents of total-isoflavones, daidzein, and genistein were at the highest value. Conclusion: The secondary fermentation process is an important link in fermentation processing of fermented soybeans and the key factor to affect the change of main chemical composition content and the finished product properties of fermented soybeans. The fermentation processing technology of fermented soybeans after optimization would lay the foundation of the regulation of production and the research of fermentation processing mechanism.

DesinFix TM 135 in fermentation process for bioethanol production

Brazil has the world's largest ethanol production from sugarcane, but bacterial contamination decreases the ethanol yields. It was shown that the biocide DesinFixTM 135 can reduce the contamination without decreasing the yeasts' viability or negatively affecting the ethanol production.

Optimization of fermentation process for metabolic engineered recombinant bacteria with methylenetetrahydrofolate reductase gene / 中国药学杂志

OBJECTIVE: The fermentation process for the metabolic engineered recombinant bacteria with methylenetetrahydrofolate reductase gene was optimized to improve L-5-methyltetrahydrofolate accumulation in the engineering bacteria. METHODS: Single- factor and orthogonal experiments were employed to investigate the optimum fermentation process of the bacteria. RESULTS: The optimum fermentation process of the bacteria was as following: sucrose was 3%, yeast extract was 1.0%, inorganic salts were NaCl-K2HPO4-MgSO4 = 1.26%:0.42%:0.02%, ammonium dihydrogen phosphate was 1.0%, inoculum amount was 5%, initial pH value was 7.5, induction time point was 2.5 h, inducer concentration was 1.4 mmol·L-1, incubation temperature was 37°C, induction duration was 6 h, addition degree of inducer was 2. Under the optimum fermentation process, the optimization results were as following: the bacteria amount had a 36.4% enhancement, the specific activity of methylenetetrahydrofolate reductase 46.8%, the L-5-methyltetrahydrofolate accumulation 66.1%, comparing with the original fermentation process. CONCLUSION: The research work provides experimental evidence for amplifying of the bacteria in agitating fermentor.

A Rapid Filtration Sampling Probe for Fermentation Broth On-line Measurement / 分析化学

An inner-bioreactor rapid filtration sampling probe was designed to resolve the problems of continuous sampling the fermentation broth in on-line measurement the fermentation process. The probe consists of a support, a plug valve, an O-ring, a filter membrane and a pipe cover. It can be inserted directly into the bio-reactor withstanding high-temperature sterilization before fermentation, or preventing the bacteria's invasion during sampling. Fermentation broth in tank is cross-flow filtrated by a cylindrical ceramic membrane which is used as filtration membrane, and then the sterile permeated liquid is transported to the analyzer by a peristaltic pump. The sampling experiments using this device was described for sampling glucose solution indicating it is suitable for fermentation online sampling. The sampling rate is up to 3. 0 mL/ min, glucose permeability maintains 100% , and the delay time caused by the probe is about 2 min.

Optimization of Fermentation Process of Compound Chinese Medicine by Response Surface Methodology / 中国中医药信息杂志

Objective To investigate the factors that influence the fermentation process of compound Chinese medicine and determining the optimum fermentation with single factor experiment and response surface methodology. Methods Through controlling the factors in the fermentation process of compound Chinese medicine (such as fermentation bacteria, fermentation time, fermentation temperature, inoculum amount, etc.), with its increase rate of total peak area as evaluation indicator, the alcohol extracts before and after fermentation were monitored and comparative evaluated by HPLC, and the optimum fermentation process was determined by response surface methodology. Results The fermentation process optimized by single factor experiment and response surface methodology was as follows:SZ-2 strain served as the fermentation bacteria, temperature was 33 ℃, inoculum amount was 4%, and time was 3.5 d, the average increase rate of total peak area was 31.24%. Conclusion HPLC can be used to identify and evaluate the fermentation of compound Chinese medicine under the different factors, and to clarify the optimal fermentation process by response surface methodology, which provide reference for the development of fermentation process.

Inoculum padronization for the production of cutinase by Fusarium oxysporum

Cutinase is a versatile enzyme showing several interesting properties for application in industrial processes. The widespread use of this enzyme depends on the development of an efficient and low-cost production system. One of the most important steps in a fermentation process is the standardization of the inoculum characteristics. In this study, the production of cutinase by Fusarium oxysporum showed a statistically significant relationship with both the inoculum size and the inoculum PDA pH. The greatest activities were 19.1 U/mL at PDA pH 7.0 and 22.72 U/mL using an aliquot of 12.72 x 10(7) spores/mL. The macroscopic characteristics of the colonies of Fusarium oxysporum changed according to the variation of the medium pH, with the best results recorded in those colonies presenting a cotton white aspect.

Cutinase é uma enzima versátil, que apresenta propriedades interessantes para aplicação em processos industriais. O uso desta enzima em larga escala depende do desenvolvimento de um sistema de produção eficiente e de baixo custo. Uma das etapas mais importantes em um processo de fermentação é a padronização do inóculo. Neste estudo, houve uma associação estatisticamente significativa entre a produção de cutinase por Fusarium oxysporum e tamanho do inóculo e pH do meio PDA. As maiores atividades de cutinase foram 19,1 U/mL em PDA com pH 7,0 e 22,72 U/mL empregando um inóculo de 12,72 x 10(7) esporos/mL. As características macroscópicas das colônias de Fusarium oxysporum mostraram alterações em função do pH do meio, com as maiores atividades sendo registradas em presença de colônias brancas com aspecto cotonoso.

High Density Fermentation Control of Pichia pastoris / 中国生物工程杂志

One of the important strategies for improving recombinant protein expression in Pichia pastoris is optimizing control during the high density fermentation process. Some fermentation control has been used and got obviously higher expression level:optimization of basal salt medium composition; using two stage pH and two stage temperature control during cell growth and induction stage; appropriately increasing dissolved oxygen; selecting optimal cell density prior to induction (CWPTI) and specific growth rate prior to induction (?PTI),using limit amount of initial glycerolin the basic media and feeding it during the fermentation with exponential fed-batch Model,selecting appropriate methanol feeding strategy by methanol-limited feeding batch (MLFB),oxygen-limited feeding batch (OLFB),methanol-unlimited feeding batch (MNLFB) or temperature-limited feeding batch (TLFB). Some research suggested that the expression level of the recombinant protein increase 9 times after optimizing the fermentation control.