Bacillus subtilis var. amyloliquefaciens biosynthesis of extracellular protease possessing coagulase activity and formed under conditions of limiting the nitrogen sources in the medium

An exocellular protease having the activity of coagulase was synthesized by Bacillus subtilis var. amyloliquefaciens when the growth medium contained no nitrogen sources. The removal of a nitrogen source from the medium was found to induce the synthesis of exoproteases by washed bacterial cells. Protein as a sole source of nitrogen in the medium inhibited rather than induced the biosynthesis of proteases possessing the activity of coagulase by the cells. The production of exoproteases with the coagulase activity was inhibited when a mixture of amino acids was used as a sole nitrogen source.     

Production of lipase free of citrinin by Penicillium citrinum

Lipase (Glycerol ester hydrolase E.G. 3.1.1.3) from a Brazilian strain of Penicillium citrinum free of the mycotoxin citrinin has been investigated. Citrinin production was inhibited by using culture medium containing olive oil, soybean oil and corn oil as carbon sources. Potassium concentration and pH play an important role in citrinin production. Potassium concentration lower than 30 mM and pH below 4.5 inhibited the mycotoxin production. P. citrinum produced lipase free of extraneous proteins and citrinin when cultured using, as nitrogen source, ammonium sulphate (lipase activity of 7.88 U/mg) and yeast extract (lipase activity of 4.95 U/mg) with olive oil as carbon source. This data is relevant to the larger scale production of lipases for food technology applications, from Penicillium citrinum.     

溶剂稳定性蛋白酶产生菌Bacillus licheniformis YP1的发酵优化

溶剂稳定性蛋白酶产生菌Bacillus licheniformis YP1分离自油田土样。考察了碳源、氮源、金属离子等营养因素对YP1菌株发酵产溶剂稳定性蛋白酶的影响。YP1菌株发酵产胞外蛋白酶的最佳碳源为淀粉,果糖、甘露糖和乳糖显著抑制产酶;最佳氮源为酵母膏,干酪素、酵母粉和牛肉膏促进产酶,玉米浆和尿素显著抑制产酶。Mn^2＋可以显著促进酶活,Mg^2＋可以促进产酶,在初步优化的培养条件下,YP1菌株的胞外蛋白酶产量达980U。     

Extracellular expression of pullulanase from Bacillus naganoensis in Escherichia coli

The pullulanase encoding gene from Bacillus naganoensis was successfully overexpressed in Escherichia coli both intracellularly and extracellularly using expression vector pET22b (+). The distribution of recombinant protein was significantly affected by temperature and carbon and nitrogen sources. The highest levels of extracellular and intracellular production of the target protein were observed at 25 and 20 °C, respectively. The addition of maltose, dextrin, pullulan, and soluble starch to the culture medium caused significant increases in the extracellular yield of pullulanase, while glucose strongly inhibited pullulanase production. The results show that the optimal conditions for maximum yield of extracellular pullulanase required high levels of carbon source and a limited nitrogen supply, while low concentrations of carbon and nitrogen source favored intracellular pullulanase expression. High concentrations of nitrogen source strongly inhibited the production of pullulanase.     

Effects of nitrogen sources on production and composition of sophorolipids by <Emphasis Type="Italic">Wickerhamiella domercqiae</Emphasis> var. <Emphasis Type="Italic">sophorolipid</Emphasis> CGMCC 1576

The effects of nitrogen sources on growth of sophorolipid-producing yeast, Wickerhamiella domercqiae var. sophorolipid CGMCC 1576 and on production and composition of sophorolipids were studied. Organic nitrogen sources are more favorable for accumulation of biomass than inorganic ones. Presence of ammonium ion from different inorganic nitrogen sources (except NH₄HCO₃) greatly inhibited the production of lactonic sophorolipids. However, when organic nitrogen sources were used, lactonic sophorolipid production was strongly increased. Production of crystalline lactonic sophorolipids from organic/inorganic nitrogen sources was enhanced with the increase of pH value adjusted by sodium hydroxide or sodium citrate solution. Fourier-transform infrared (FT-IR), gas chromatography mass spectrometry (GC-MS), high-performance liquid chromatography (HPLC), and mass spectra (MS) were employed to compare the composition of sophorolipid mixture obtained from different nitrogen sources. More than 15 acidic sophorolipid molecules and only 4 lactonic sophorolipid molecules were produced by using 1.27 g/l ammonium sulfate as nitrogen source; they were separated by preparative HPLC and their structures were elucidated by MS. These results suggest extraordinary regulatory effects of nitrogen source on growth and sophorolipid synthesis of W. domercqiae var. sophorolipid.     

正交实验优选光合细菌混合菌群产氢的最佳条件

对光合细菌混合菌群产氢影响因子进行了实验研究。通过单因素实验和正交实验, 系统考察了碳源、氮源、碳源浓度、氮源浓度、初始pH值、光照方式、接种量等因素对产氢量的影响, 实验得出最佳工艺条件为: 采用3号菌群, 碳源为葡萄糖, 碳源浓度为3 g/L, 氮源为尿素, 氮源浓度为9 g/L, 接种量为10%, pH值为8.5, 光照方式为12 h光照-12 h黑暗交替光照, 培养温度为30°C。菌种、碳源、碳源浓度、氮源是影响产氢量的重要因素。     

Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3

A thermophilic isolate Bacillus coagulans BTS-3 produced an extracellular alkaline lipase, the production of which was substantially enhanced when the type of carbon source, nitrogen source, and the initial pH of culture medium were consecutively optimized. Lipase activity 1.16 U/ml of culture medium was obtained in 48 h at 55 degrees C and pH 8.5 with refined mustard oil as carbon source and a combination of peptone and yeast extract (1:1) as nitrogen sources. The enzyme was purified 40-fold to homogeneity by ammonium sulfate precipitation and DEAE-Sepharose column chromatography. Its molecular weight was 31 kDa on SDS-PAGE. The enzyme showed maximum activity at 55 degrees C and pH 8.5, and was stable between pH 8.0 and 10.5 and at temperatures up to 70 degrees C. The enzyme was found to be inhibited by Al3+, Co2+, Mn2+, and Zn2+ ions while K+, Fe3+, Hg2+, and Mg2+ ions enhanced the enzyme activity; Na+ ions have no effect on enzyme activity. The purified lipase showed a variable specificity/hydrolytic activity towards various 4-nitrophenyl esters.     

Bacteriocin production with Lactobacillus amylovorus DCE 471 is improved and stabilized by fed-batch fermentation

Amylovorin L471 is a small, heat-stable, and hydrophobic bacteriocin produced by Lactobacillus amylovorus DCE 471. The nutritional requirements for amylovorin L471 production were studied with fed-batch fermentations. A twofold increase in bacteriocin titer was obtained when substrate addition was controlled by the acidification rate of the culture, compared with the titers reached with constant substrate addition or pH-controlled batch cultures carried out under the same conditions. An interesting feature of fed-batch cultures observed under certain culture conditions (constant feed rate) is the apparent stabilization of bacteriocin activity after obtaining maximum production. Finally, a mathematical model was set up to simulate cell growth, glucose and complex nitrogen source consumption, and lactic acid and bacteriocin production kinetics. The model showed that bacterial growth was dependent on both the energy and the complex nitrogen source. Bacteriocin production was growth associated, with a simultaneous bacteriocin adsorption on the producer cells dependent on the lactic acid accumulated and hence the viability of the cells. Both bacteriocin production and adsorption were inhibited by high concentrations of the complex nitrogen source.     

Production and partial characterization of high molecular weight extracellular alpha-amylase from Thermoactinomyces vulgaris isolated from Egyptian soil

Optimizing production of alpha-amylase production by Thermoactinomyces vulgaris isolated from Egyptian soil was studied. The optimum incubation period, temperature and initial pH of medium for organism growth and enzyme yield were around 24 h, 55 degrees C and 7.0, respectively. Maximum alpha-amylase activity was observed in a medium containing starch as carbon source. The other tested carbohydrates (cellulose, glucose, galactose, xylose, arabinose, lactose and maltose) inhibited the enzyme production. Adding tryptone as a nitrogen source exhibited a maximum activity of alpha-amylase. Bactopeptone and yeast extract gave also high activity comparing to the other nitrogen sources (NH4CI, NH4NO3, NaNO3, KNO3, CH3CO2NH4). Electrophoresis profile of the produced two alpha-amylase isozymes indicated that the same pattern at about 135-145 kDa under different conditions. The optimum pH and temperature of the enzyme activity were 8.0 and 60 degrees C, respectively and enzyme was stable at 50 degrees C over 6 hours. The enzyme was significantly inhibited by the addition of metal ions (Na+, Co2+ and Ca2+) whereas CI- seemed to act as activator. The enzyme was not affected by 0.1 mM EDTA while higher concentration (10 mM EDTA) totally inactivated the enzyme.     

沼泽红假单胞菌乙酸光合放氢研究

依据光合细菌生长代谢特性和有机废水降解主要产物类型,11种有机物被用于沼泽红假单胞菌（Rhodopseudomonas palustris）Z菌株的光合产氢研究,其中,乙酸反应体系产氢活性最高。在此基础上,研究了该菌株的生长与产氢动力学行为,探求了影响该菌株光合放氢的主要限制性影响因素。结果表明,该菌株产氢与生长部分相关。种子培养基和菌龄对产氢活性有明显影响。细胞最适产氢和生长所需要的光照强度和温度基本一致。当种子来源于硫酸铵高菌龄预培养物或谷氨酸钠对数期预培养物时,该菌株产氢活性显著增加,产氢延滞期明显缩短。氧浓度和接种量对产氢活性也有显著影响。供氢体和氮源浓度直接决定细胞的生长与光放氢活性。在低于70 mmol/L乙酸钠和15 mmol/L谷氨酸钠时,产氢活性随底物浓度的增加而增强。谷氨酸钠浓度高于15mmol/L时,由于游离NH₄⁺的出现,产氢活性受到抑制,但却明显刺激细胞的生长。在标准状况下,该菌株的最大产氢速率可达19.4 mL·L^-1·h^-1。     

Effect of levulinic acid on pigment biosynthesis in Agmenellum quadruplicatum.

When levulinic acid was added to a growing culture of the cyanobacterium (blue-green alga) Agmenellum quadruplicatum PR-6, delta-aminoelevulinic acid accumulated in the medium and chlorophyll a synthesis and cell growth were inhibited, but there was a small amount of c-phycocyanin synthesis. The amount of delta-aminolevulinic acid produced in the treated culture did not fully account for the amount of pigment synthesized in the untreated control. Levulinic acid and either sodium nitrate or ammonium chloride were added to nitrogen-starved cultures of PR-6, and delta-aminolevulinic acid production and chlorophyll a and c-phycocyanin content were monitored. When ammonium chloride was added as a nitrogen source after nitrogen starvation, the cells recovered more rapidly than when sodium nitrate was added as a nitrogen source. In cultures recovering from nitrogen starvation, synthesis of c-phycocyanin occurred before synthesis of chlorophyll a.     

Regulation of biosynthesis of bacilysin byBacillus subtilis

Summary Production of the dipeptide antibiotic bacilysin byBacillus subtilis 168 was growth associated and showed no evidence of repression by glucose or sucrose. Carbohydrates other than glucose and sucrose yielded lower specific titers of bacilysin. Bacilysin production in three such carbon sources (maltose, xylose, ribose) was delayed until growth slowed down. Ammonium salts were poor for bacilysin production when used as the sole nitrogen source. When added to the standard medium containing glutamate, they suppressed antibiotic production. Aspartate was slightly better than glutamate for antibiotic production as sole nitrogen source. No other nitrogen source tested, including inorganic, organic or complex, approached the activity of glutamate or aspartate. When added to glutamate, casamino acids, phenylalanine and alanine (a substrate of bacilysin synthetase) suppressed bacilysin production while stimulating growth. Phosphate provided for optimum growth and production at 7.5 mM and both processes were inhibited at higher concentrations. Ferric citrate stimulated growth and inhibited bacilysin production, the effects being due to both the iron and the citrate components. Elimination of ferric citrate stimulated production as did increasing the concentration of Mn to its optimum concentration of 6.6×10^–4M.     

Lysine catabolism inStreptomyces ambofaciens producer of macrolide antibiotic,spiramycin

Spiramycin production was highly stimulated when lysine was used as the sole nitrogen source. This amino acid was catabolized by the -transaminase pathway characterized by dosage of cadaverine aminotransferase (CAT) enzyme. The Km_cadaverine was of 57mM. CAT was highly induced by lysine (634% in comparison with ammonium). Addition of 40mm of ammonium in a culture begun with 20mm of lysine as the sole initial nitrogen source repressed CAT biosynthesis by 24% but did not affect spiramycin production seriously. Addition of 20mm of lysine in a culture started with 40mm ammonium induced CAT biosynthesis of 425%, but did not allow spiramycin production. In these two cases, spiramycin production seems to be conditioned by the nitrogen source initially present in the culture medium. CAT activity was inhibited by ammonium ions (33% at 20mm), whereas lysine had no effects.     

Involvement of lignin peroxidase in the decolourization of black olive mill wastewaters by Geotrichum candidum

AIM: Decolourization of black olive mill wastewaters (OMW) by depolymerization of phenolic compounds by Geotrichum candidum. METHODS AND RESULTS: Our results show that G. candidum is able to grow on black OMW supplemented with carbon source and nitrogen. The Geotrichum growth decreased the pH and induced a 49% of colour removal when the black OMW was supplemented with glycerol and diammonium tartrate (20 mm ammonium). An improvement of 10% of colour removal was observed when the culture was supplemented with veratryl alcohol. The decolourization was inhibited with glutamate as nitrogen source. CONCLUSION: Our results suggest the potential use of G. candidum in black OMW decolourization and support the concept that lignin peroxidase (LiP) of G. candidum is involved in the depolymerization of phenolic compounds. Significance and Impact of the Study: This is the first report of LiP production by G. candidum on OMW.     

Bacteriocin Production with Lactobacillus amylovorus DCE 471 Is Improved and Stabilized by Fed-Batch Fermentation

Amylovorin L471 is a small, heat-stable, and hydrophobic bacteriocin produced by Lactobacillus amylovorus DCE 471. The nutritional requirements for amylovorin L471 production were studied with fed-batch fermentations. A twofold increase in bacteriocin titer was obtained when substrate addition was controlled by the acidification rate of the culture, compared with the titers reached with constant substrate addition or pH-controlled batch cultures carried out under the same conditions. An interesting feature of fed-batch cultures observed under certain culture conditions (constant feed rate) is the apparent stabilization of bacteriocin activity after obtaining maximum production. Finally, a mathematical model was set up to simulate cell growth, glucose and complex nitrogen source consumption, and lactic acid and bacteriocin production kinetics. The model showed that bacterial growth was dependent on both the energy and the complex nitrogen source. Bacteriocin production was growth associated, with a simultaneous bacteriocin adsorption on the producer cells dependent on the lactic acid accumulated and hence the viability of the cells. Both bacteriocin production and adsorption were inhibited by high concentrations of the complex nitrogen source.     

Bioproduction of riboflavin from molasses and lentils

Fermentation studies were carried out for the bioproduction of riboflavin with an agroindustrial byproduct, molasses as the carbon source and lentils as the nitrogen source using E. ashbyii strain. With the previously recommended 1.5% (w/v) molasses, lentils at a concentration of 3% (w/v) was found to be the optimum. Acidic medium was found to be favorable for riboflavin production when the molasses to lentils ratio was greater than one and neutral medium when the ratio was one. Effect of agitation on vitamin production was also studied and it was observed that 300 rpm gives a higher yield of product.     

The Influence of n-Propanol on the Growth and Conidiation of Pestalotia rhododendri

Pestalotia rhododendri was exposed to vapours from 1 ml propanol solution in water and linear growth, formation of aerial hyphae and production of conidia were determined. A special Petri dish technique was used and maximum stimulation of conidial formation was induced by the vapours from a propanol concentration of 3–4 % (v/v) at 25°C. When propanol was added directly to the medium, a concentration of 1.2 × 10^?2M was optimal for growth and sporulation at 30°C. Sporulation stimulated by propanol was observed at temperatures from 20–32°C, with an optimum at 30°C. Certain observations indicated that an exposure to propanol for 24 hours was enough to induce a stimulated spore production. The stimulation was noticed on different media at 25°C, and was more pronounced at 30°C. One exception was observed. Propanol did not promote sporulation when the fungus was grown on maltagar at 30°C. Propanol 3 ° (v/v) in combination with the standard medium containing (NH₄)₂-tartrate as sole nitrogen source, inhibited the linear growth at 15–20°C, was inactive at 22.5° and 25°C, and stimulated growth at 27.5–31°C. The stimulatory effect was maximal at 30°C. Other media were tested at 25° and 30°C. At both temperatures stimulations of linear growth caused by propanol were observed with a medium containing KNO₃ as sole nitrogen source, and inhibitions with maltagar and another medium containing l -asparaginc as sole nitrogen source. The linear growth could be either inhibited or stimulated while the sporulation was stimulated.     

碳源、氮源对异养单针藻Monoraphidium sp. FXY-10油脂积累和脂肪酸组成的影响

研究了碳源与氮源对单针藻Monoraphidium sp. FXY-10异养培养的影响。以BG-11为基础培养基,通过添加不同类型、浓度梯度碳源和氮源,比较分析微藻生物量、油脂积累以及脂肪酸组成。结果表明,以葡萄糖作碳源,硝酸钠为氮源,微藻细胞积累的油脂是理想的生物柴油制备原料。硝酸钠浓度分别为1.00、3.00和5.00 g/L时,对油脂产量影响不显著(P>0.05)。葡萄糖浓度为10.00 g/L,硝酸钠为氮源油脂产量达到实验最高值0.84 g/L,其油脂脂肪酸组成主要由C16:0和C18:1等短链饱和脂肪酸和单不饱和脂肪酸组成,不饱和度值(DU)为61.98,相对偏低。     

影响白腐真菌AH28-2菌株漆酶合成的因子和发酵条件

White-rot fungus AH28-2, a newly isolated strain, produced effectively laccase by induction when grown on a synthetic medium. Aromatic compounds of low molecular weight had an inducing influence on laccase production and its isoenzyme compositions. The using of o-toluidine or syringic acid had the best inducing effect. Cu2+ concentration in medium had distinguished effect on laccase production. Enzyme activity was notably increased by Cu2+ and reached the maximum when Cu2+ final concentration was 5 mumol/L. Mn2+ inhibited the synthesis of laccase. Carbon and nitrogen limitation were not beneficial to laccase synthesis, while high nutrient organic medium was beneficial to the growth of cell and the synthesis of laccase. Using cellobiose as the sole carbon source, the highest level enzyme activity reached 82,923. 7 u/L under the condition of optimum fermentation with ABTS as substrate. This enzyme activity was 2.9-fold higher compared to the reported data on international references in recent years.     

毛栓菌(Tramets trogii)液体培养产漆酶的研究

实验研究了碳源、氮源和综合因素对毛栓菌(Tramets trogii)液体培养产漆酶和生长情况的影响。碳源为玉米面、氮源为豆饼粉时,菌株的产漆酶酶活最高;碳源为可溶性淀粉,氮源为麦麸时,对菌株的生长有利;在液体条件下,菌株产漆酶的最佳培养基组成为玉米面3.5%,豆饼粉4.0%,KH2PO40.3%,MgSO40.15%,VB10.04%。