The 4‐acetylantroquinonol B isolated from mycelium of Antrodia cinnamomea inhibits proliferation of hepatoma cells

BACKGROUND: Antrodia cinnamomea is known for its antihepatoma activity, yet the identity of its active compound was unclear. In this study, a 5‐ton fermenter was used to prepare sufficient mycelium of A. cinnamomea for active compound isolation and identification. RESULTS: Using antiproliferative activity toward HepG2 cells as guidance in the isolation process, 4‐acetylantroquinonol B was purified and identified to be the major bioactive compound of A. cinnamomea cultivated by submerged fermentation. The median effective doses (EC₅₀) of 4‐acetylantroquinonol B for HepG2 cells were 0.10 ± 0.00 and 0.08 ± 0.00 µg mL^?1 for 72 and 96 h treatments, respectively. The selective indices of 4‐acetylantroquinonol B were 100 and 125 for 72 and 96 h treatments, respectively, indicating that this compound had high selective activity for hepatoma cells. CONCLUSION: 4‐Acetylantroquinonol B is the major antihepatoma constituent of Antrodia cinnamomea mycelium produced by submerged fermentation. Copyright © 2010 Society of Chemical Industry     

Screening of medium composition for the free radical‐scavenging properties by Antrodia cinnamomea

The effects of nutritional conditions on the scavenging capacity on superoxide anion radical and 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) free radical of ethanolic extracts from filtrates of Antrodia cinnamomea were investigated in shake‐flask cultures. Culture medium significantly affected inhibition of superoxide anion generation and DPPH free radical scavenging activity. In contrast, both scavenging capabilities on superoxide anion radical and DPPH free radical were found to be greatly affected by varying the carbon source, the nitrogen source, the growth factors or the trace elements. The maximal inhibition of superoxide anion generation could be obtained when the culture medium compositions were: xylose, ammonium oxalate, nicotinic acid, NaH₂PO₄ and CuSO₄; while a maximal DPPH free radical scavenging activity could be achieved when the medium composition setting as: fructose, peptone, nicotinic acid, KH₂PO₄ and CaCl₂ (the scavenging effects on superoxide anion radical and DPPH free radical were increased to 96.3 ± 0.3% and 29.0 ± 1.0%, respectively). We proved that controlling the culturing conditions and modifying the composition of the medium can dramatically enhance the scavenging ability on superoxide anion radical and DPPH free radical of A. cinnamomea.     

Comparison of the performances of different fermentation strategies on cell growth and bacteriocin production by Lactobacillus curvatus CWBI‐B28

The dynamics of cell growth and bacteriocin production by Lactobacillus curvatus CWBI‐B28 in modified De Man/Rogosa/Sharp (mMRS) broth with various concentrations of glucose and complex nitrogen source (CNS; peptone, yeast extract and meat extract) was investigated in flask fermentations and in a laboratory fermentor using batch and fed‐batch cultivations. In fed‐batch fermentation the rate of feeding of the reactor with the substrates was either maintained constant (0.12 L h^?1) or varied exponentially as a function of time. The results showed that both cell growth and bacteriocin activity were influenced by changes in the concentrations of glucose and CNS. Optimal growth and bacteriocin activity were obtained in mMRS broth containing 40 g L^?1 glucose and 40 g L^?1 CNS (mMRS_40/40). A bacteriocin titre of 4266 AU mL^?1 and a cell count of 8.7 log colony‐forming units (cfu) mL^?1 were recorded when this medium was used for cultivation. In batch fermentation using the same medium, a higher cell count (9.5 log cfu mL^?1) and twice as much bacteriocin as in flask fermentation were produced. The highest bacteriocin titre (8533 AU mL^?1) was obtained with fed‐batch fermentation at an exponentially varying rate of feeding. Bacteriocin activity and cell dry mass did not always correlate. Copyright © 2007 Society of Chemical Industry     

樟芝深层发酵培养条件的优化

本文对樟芝深层发酵条件进行了详细的研究,确定了樟芝发酵的最优方案是:2%麸皮,0.2%蛋白胨,0.3%磷酸氢二钾,0.05%硫酸镁,VB1 10mg/100ml,自然pH,接种量为20%,摇瓶的装液量为120ml/250ml三角瓶,150r/min,25℃恒温培养5d,菌丝干重达到1.0g/100ml发酵液。并通过对樟芝深层培养菌丝体及发酵滤过液多糖中氨基酸营养成分及多糖含量进行了分析。     

Ethanolic extracts of Antrodia cinnamomea mycelia fermented at varied times and scales have differential effects on hepatoma cells and normal primary hepatocytes

Mycelia of Antrodia cinnamomea (AC), an edible fungus native to Taiwan, were produced by submerged fermentation with various fermentation times in 250 mL, 5 and 500 L fermentors and were evaluated for the effect of fermentation products on the viabilities of Hep3B and HepG2 hepatoma cells and normal primary rat hepatocytes. The results showed that the ethanolic extracts of AC mycelia (from 250 mL fermentation for 8 wk and 5 and 500 L fermentations for 4 wk) possessed high antihepatoma activity. The IC(50) of ethanolic extract of AC mycelia fermented for 8 wk in a 250 mL fermentor against Hep3B and HepG2 cells were 82.9 and 54.2 microg/mL, respectively. Furthermore, the IC(50) for Hep3B and HepG2, treated with ethanolic extract of AC mycelia fermented for 4 wk in the 5 L fermentor were 48.7 and 3.8 microg/mL, respectively. Those treated with ethanolic extract of AC mycelia fermented for 4 wk in the 500 L fermentor were 36.9 and 3.1 microg/mL, respectively. No adverse effects of all samples on normal primary rat hepatocytes were observed.     

Anti‐inflammatory and cytotoxic effects of ginseng extract bioconverted by Leuconostoc mesenteroides KCCM 12010P isolated from kimchi

A bioconversion technique using microorganisms has been applied to ginseng to increase content of bioactive ginsenoside and biofunctionality such as anticancer, anti‐obesity and antioxidant activities. The objective of this study was to screen lactic acid bacteria for bioconversion of ginsenosides and to evaluate anti‐inflammatory and cytotoxic effects of bioconverted ginseng extract. Strains isolated from kimchi were screened for their β‐glucosidase activities using esculin agar. Selected strain was identified based on 16S rRNA sequencing and carbohydrate fermentation. During ginseng fermentation, viable cell number and pH were determined. Bioconverted ginsenosides were analysed by HPLC. Anti‐inflammatory effects were evaluated using RAW 264.7 cells, and cytotoxic effects were determined by MTT assay. Among 166 isolates screened, Leuconostoc mesenteroides was selected for ginseng bioconversion, as it showed a higher β‐glucosidase activity and viable cell number than any of the other tested strains. After fermentation for 2 days, viable cell number was 8.8 log CFU mL^?1 and final pH was 4.8. Ginsenoside Rb₂ was bioconverted into ginsenoside Rg₃ (Rb₂ → Rd → Rg₃) by L. mesenteroides. The nitric oxide contents of 2‐day‐fermented extract decreased by as much as 25%, compared to a non‐fermented extract. The cell viabilities of HepG2, HT‐29, HeLa and LoVo treated with fermented ginseng extract also decreased by 49.7%, 20.2%, 21.0% and 8.7%, respectively, compared to those of control cells treated with non‐fermented extract. Ginseng extract bioconverted by L. mesenteroides showed anti‐inflammatory and anticancer effects. Therefore, bioconverted ginseng extract might have applications in the pharmaceutical and/or functional food industry.     

In vitro anti‐HSV‐2 activity and mechanism of action of proanthocyanidin A‐1 from Vaccinium vitis‐idaea

The aim of this study was to investigate the in vitro anti‐HSV‐2 activity and mechanism of action of proanthocyanidin A‐1, a compound isolated from Vaccinium vitis‐idaea Linn (Ericaceae). The results demonstrated that proanthocyanidin A‐1 exhibited anti‐HSV‐2 activity. The IC₅₀ value for the XTT assay was 73.3 ± 14.5 µM and the IC₅₀ and IC₉₀ values for the plaque reduction assay were 41.9 ± 2.0 and 62.8 ± 6.3 µM respectively. Proanthocyanidin A‐1 showed no cell cytotoxic effect at concentrations that blocked HSV‐2 infection, with a CC₅₀ value of 282.1 ± 27.5 µM . The mechanism studies demonstrated that proanthocyanidin A‐1 did not reduce viral infectivity but inhibited viral attachment and penetration and affected the late stage(s) of HSV‐2 infection. It was concluded that proanthocyanidin A‐1 suppressed HSV‐2 infection through many modes of action and thus merits further investigation. Copyright © 2004 Society of Chemical Industry     

Anti‐inflammatory properties of high pressure‐assisted extracts of Grifola frondosa in lipopolysaccharide‐activated RAW 264.7 macrophages

The aim of this study was to determine the in vitro anti‐inflammatory properties of the shake extract (SE) and the high pressure‐assisted extract (PE) of the mycelia of Grifola frondosa in a lipopolysaccharide‐stimulated RAW 264.7 macrophage model. The content of total polysaccharides and β‐glucans of PE at 600 MPa (PE‐600) was 41.2 and 6.2 mg g^?1 dry weight, respectively, which were significantly higher than SE extracts. The results showed that treatment with 500 μg mL^?1 of PE by 600 MPa (PE‐600) did not reduce RAW 264.7 cell viability but did significantly inhibit the production of LPS‐induced NO, PGE₂ and intracellular ROS. The PE‐600 inhibited the activation of NF‐kB and then reduced the production of LPS‐induced TNF‐α, IL‐6 and IL‐1β in a dose‐dependent manner. Thus, the PE could be used as an alternative extraction method for improving the extraction efficacy of G. frondosa and serve as an alternative source of anti‐inflammatory agents.     

Ethanol production from carob pods by Saccharomyces cerevisiae

The production of ethanol from carob pods extract by Saccharomyces cerevisiae in static and shake flask fermentation was investigated. Shake flask fermentation proved to be a better fermentation system for the production of ethanol than static fermentation. The external addition of nutrients into the carob pods extract did not improve the production of ethanol. The maximum concentration of ethanol (75 g/l) was obtained at an inoculum amount of 0.3%, a pH of 4.5, 30°C and an initial sugar concentration of 200 g/1. Under the same fermentation conditions both sterilized and non‐sterilized carob pods extract gave the same final ethanol concentration.     

响应面法优化根霉菌产麦角固醇的液体发酵工艺

以米根霉(Rhizopus oryzae ZW017)发酵产麦角固醇的产量为响应值,对其液体发酵工艺进行优化。采用HPLC法检测菌株产麦角固醇含量,在单因素筛选试验基础上,以PDB液体发酵培养为基础条件,应用响应面分析法(RSM)对碳源、氮源及发酵时间进行优化。结果表明:以葡萄糖、酵母膏分别为最佳碳、氮源;最佳工艺条件为:PDB基础培养基中添加葡萄糖3g/L、酵母膏5g/L、发酵培养9.64d,麦角固醇平均产量达5761.83μg/100mL,较优化前提高了247.86%,与构建模型理论预测值(5818.39μg/100mL)相吻合,且100mL液体培养基中麦角固醇产量占菌体细胞干质量(0.36g)的1.60%。     

Fed‐batch l‐(+)‐lactic acid fermentation of brewer's spent grain hydrolysate

Brewer's spent grain (BSG) hydrolysates were used for l ‐(+)‐lactic acid (LA) fermentation by Lactobacillus rhamnosus ATCC 7469. The aim of this study was to evaluate fed‐batch LA fermentation of BSG hydrolysate with the addition of glucose, glucose and yeast extract, and wort during LA fermentation and its effect on fermentation parameters such as LA concentration, its volumetric productivity and yield, and L. rhamnosus cell viability. The highest LA yield, volumetric productivity and concentration of 93.3%, 2.0 g/L/h, and 116.1 g/L, respectively, were achieved with glucose and yeast extract addition during fermentation. In fed‐batch fermentation with glucose and yeast extract addition significantly higher LA concentration, yield and volumetric productivity (by 194.8; 2.2, and 20.7%, respectively) were achieved compared with batch fermentation. The results indicated that fed‐batch fermentation could be used to increase LA fermentation efficiency. Copyright © 2017 The Institute of Brewing & Distilling     

Growth of the Acid-Tolerant Fungus Scytalidium acidophilum as a Potential Source of Single-Cell Protein

An acid-tolerant fungus, Scytalidium acidophilum, was cultivated in peat extract for potential use as a single-cell protein (SCP) source. In shake flask fermentations with a 0.3% yeast extract diluted peat extract medium, it was found that the best conditions for growth occurred at pH 2.0 and 25°C. The maximum biomass concentration was produced after 10 days, with no significant differences for agitation rates between 100 and 200 rpm. Microbial contamination was not observed in nonaseptic operations. The protein content and amino acid composition of the mycelium produced compares with those reported for S. acidophilum grown in other fermentation media. The advantages of growing S. acidophilum in acid peat extract are presented.     

Influence of degree of hydrolysis on functional properties and angiotensin I‐converting enzyme‐inhibitory activity of protein hydrolysates from cuttlefish (Sepia officinalis) by‐products

BACKGROUND: In Tunisia the cuttlefish‐processing industry generates large amounts of solid wastes. These wastes, which may represent 35% of the original material and constitute an important source of proteins, are discarded without any attempt at recovery. This paper describes some functional properties and the angiotensin I‐converting enzyme (ACE)‐inhibitory activity of protein hydrolysates prepared by hydrolysis of cuttlefish (Sepia officinalis) by‐products with crude enzyme extract from Bacillus licheniformis NH1. RESULTS: Cuttlefish by‐product protein hydrolysates (CPHs) with different degrees of hydrolysis (DH 5, 10 and 13.5%) were prepared. All CPHs contained 750–790 g kg^?1 proteins. Solubility, emulsifying capacity and water‐holding capacity increased while fat absorption and foaming capacity decreased with increasing DH. All hydrolysates showed greater fat absorption than the water‐soluble fraction from undigested cuttlefish by‐product proteins and casein. CPHs were also analysed for their ACE‐inhibitory activity. CPH3 (DH 13.5%) displayed the highest ACE inhibition (79%), with an IC₅₀ value of 1 mg mL^?1. CONCLUSION: Hydrolysis of cuttlefish by‐product proteins with alkaline proteases from B. licheniformis resulted in a product with excellent solubility over a wide pH range and high ACE‐inhibitory activity. This study suggests that CPHs could be utilised to develop functional foods for prevention of hypertension. Copyright © 2010 Society of Chemical Industry     

纳他霉素高产菌株发酵培养基优化

考察不同碳源、氮源及前体物质对菌株Streptomyce natalensis摇瓶发酵生产纳他霉素的影响。试验表明,纳他霉素摇瓶发酵培养基最佳碳源为葡萄糖、糊精,浓度分别为40g／L和32g／L;最佳氮源为酵母粉,浓度为4．5g／L;前体物质丙酸钠加入量为0．20％,最适加入时间为对数生长后期（48h）。     

Polyphenols and phenolic acids from strawberry and apple decrease glucose uptake and transport by human intestinal Caco‐2 cells

The effect of polyphenols, phenolic acids and tannins (PPTs) from strawberry and apple on uptake and apical to basolateral transport of glucose was investigated using Caco‐2 intestinal cell monolayers. Substantial inhibition on both uptake and transport was observed by extracts from both strawberry and apple. Using sodium‐containing (glucose transporters SGLT1 and GLUT2 both active) and sodium‐free (only GLUT2 active) conditions, we show that the inhibition of GLUT2 was greater than that of SGLT1. The extracts were analyzed and some of the constituent PPTs were also tested. Quercetin‐3‐O‐rhamnoside (IC₅₀=31 μM), phloridzin (IC₅₀=146 μM), and 5‐caffeoylquinic acid (IC₅₀=2570 μM) contributed 26, 52 and 12%, respectively, to the inhibitory activity of the apple extract, whereas pelargonidin‐3‐O‐glucoside (IC₅₀=802 μM) contributed 26% to the total inhibition by the strawberry extract. For the strawberry extract, the inhibition of transport was non‐competitive based on kinetic analysis, whereas the inhibition of cellular uptake was a mixed‐type inhibition, with changes in both V_max and apparent K_m. The results in this assay show that some PPTs inhibit glucose transport from the intestinal lumen into cells and also the GLUT2‐facilitated exit on the basolateral side.     

粉被虫草液体培养条件的优化

通过摇瓶液体培养正交试验获得了粉被虫草的液体培养条件,即蔗糖2.5%,马铃薯淀粉2%,黄豆0.5%,牛肉膏0.5%,酵母膏0.1%,K2HPO4 0.1%,KCl 0.02%,MgSO4•7H2O 0.05%,pH5～7,装液量(100ml/250ml摇瓶),加15颗玻璃珠作分散剂,5%接种量,旋转式摇床150r/min,28℃培养7d,达到最大菌丝干重31.86g/L,于9d获得最大胞内产物3.13g/L。同时,对摇瓶液体培养和生物反应器液体深层分批培养的动力学做了较系统的研究,无论生物反应器还是摇瓶,其培养过程中相对应的pH、残糖、氨基氮、菌丝干重、胞内产物等参数变化趋势基本一致,但生物反应器培养时间大大缩短,48h即可达到最大菌丝干重29.97g/L,54h达到4.95g/L的最高胞内产物产率。生物反应器设定培养温度28℃、装液量65%、接种量10%、0.2%的消泡剂、搅拌转速350r/min、通气量(12±3)L/min、罐压1.1MPa、初始pH6.5,培养基配方用食用蔗糖3%、蔗糖糖蜜2%、花生0.5%分别替代蔗糖、马铃薯淀粉、牛肉膏以降低成本。实验表明,粉被虫草的液体培养条件基本能达到工业发酵水平的要求。     

The influence of tempeh fermentation and conventional cooking on anti‐nutrient level and protein bioavailability (in vitro test) of grass‐pea seeds

BACKGROUND: The solid state fermentation tempeh type is known to result in the decomposition of anti‐nutrients and the improvement of nutritional value of legume seeds. The aim of the research was to study the influence of tempeh fermentation and conventional cooking on (1) levels of 3‐N‐oxalyl‐L ‐2,3‐diaminopropionic acid (ODAP), soluble phenols, trypsin inhibitors and inositol phosphates; and (2) the in vitro bioavailability of proteins in grass‐pea seeds. RESULTS: Tempeh fermentation reduced the level of ODAP, trypsin inhibitors and phytates by 93, 99 and 22%, respectively, and increased protein bioavailability by about 25%. Protein bioavailability from conventionally cooked seeds was higher than from fermented seeds. However, during the in vitro test more soluble protein (approx. 10%) was released from 100 g tempeh product than from cooked seeds. CONCLUSIONS: Solid state fermentation and cooking resulted in seeds of comparable quality but the tempeh contained much less ODAP, which is very promising in the context of popularising grass pea. Copyright © 2008 Society of Chemical Industry     

丛梗孢酵母产赤藓糖醇的诱变选育

利用微波-硫酸二乙酯复合诱变对产赤藓糖醇丛梗孢酵母E54进行处理,以高渗平板和摇瓶发酵为筛选方法,得到遗传稳定的诱变高产株EW29;再采用氮离子注入对EW29进行诱变处理,摇瓶发酵筛选得到诱变株EN59,其90h发酵液中赤藓糖醇产量达到55.13g/L,较EW29提高20.3%,较E54提高36.9%,遗传稳定性较好。对突变株EN59的发酵培养基进行了优化,在优化培养基葡萄糖250g/L、酵母膏5g/L、KH2PO4 0.3g/L、MnSO4 ·4H2O 0.04g/L、CuSO4 ·5H2O 0.03g/L,初始pH4的条件下,90h发酵液中赤藓糖醇平均产量达到69.00g/L以上。在优化培养基的基础上进行5L罐发酵放大实验,发酵126h赤藓糖醇产量达到71.14g/L。     

On‐Line Estimation and Control of Apparent Extract Concentration in Low‐Malt Beer Fermentation

Apparent extract concentration, which is defined as the apparent total dry matter concentration in the media calculated by measurement of the specific gravity, is a very important variable in the process of beer and low‐malt beer fermentation. In the present study, ethanol formation and the residual apparent extract concentration were estimated using the CO₂ concentration in the exhaust gas. A simple mathematical model involving the CO₂ production rate, temperature and cell activity was constructed and a novel system for on‐line estimation and control of apparent extract concentration was developed. In the control system for the residual apparent extract concentration, cell activity was estimated by monitoring the differences in actual CO₂ production rate. In addition, the future trajectory of the apparent extract concentration was predicted. The degree of fermentation, i.e., the residual apparent extract concentration trajectory, was controlled by raising or lowering the temperature, taking into account the estimated cell activity at that time. The residual apparent extract concentration reached the target value at a given time, using the on‐line estimation of apparent extract concentration and temperature control.