Studies on the Lipid Production by Penicillium lilacinum

Lipid production by Penicillium lilacinum in media containing various ratios of carbon and nitrogen has been studied. Factors affecting the lipid synthesis from diverse sources of carbon and nitrogen have been observed. Out of the nitrogen sources studied sodium nitrate produced 22.5% fat. Sucrose proved to be a better source of carbon than xylose and maltose. The same yield of fat was obtained by replacing sucrose with molasses and sulphite waste liquor. Chemical examination of the fat reveals that it is composed of the usually occurring fatty acid glycerides.     

Studies on the Possibility of Producing Fats from Food Wastes by Using Micro-organisms: I: Factors Affecting Fat Production from Different Fungi

There is a great shortage of edible oils which must be compensated. Such shortage leads to the continuous research for alternative sources of edible oils which has therefore considerable national importance though little interest in exploiting micro-organisms for this purpose is evident. Factors affecting fat production from different fungi including incubation temperature, incubation period, pH-value, sugar concentration, spore count and carbon: nitrogen (C/N) ratio were studied. The optimum conditions enhancing the production of lipids from molasses media as food wastes by using Aspergillus niger, Aspergillus oryzae, Penicillium roqueforti and Rhizopus arrhizus were: 30,25,30 and 30° C for incubation temperature; 3, 7, 7 and 7 days for incubation period; 5,50:1; 7,70 :1;5,50: land 5,50 :1 for glucose percent and C/N ratio; 5,6,6 and 7 for pH-values and 2 × 10⁴,1.5 × 10⁴,2 × 10⁴ and 1.3 × 10⁵ for spore concentrations respectively. The oil contents of mycelium and fat coefficient obtained from the studied fungi grown in carrot wastes media were less than that resulted in molasses media.     

Efficient production of l-lactic acid from chicken feather protein hydrolysate and sugar beet molasses by the newly isolated Rhizopus oryzae TS-61

The aim of this study was to investigate production of l-lactic acid from molasses and chicken feather protein hydrolysate (CFP) by the newly isolated Rhizopus oryzae TS-61. R. oryzae TS-61 was capable of utilizing molasses sucrose and CFP as carbon and nitrogen sources, respectively. In contrast to yeast extract and ammonium sulfate, CFP had potential not only to prevent excessive pH changes and foaming but also to provide smaller uniform pellet formation in during fermentation. Thanks to these properties, it was concluded that CFP might have resulted in higher l-lactic acid production than the other two nitrogen sources (yeast extract and ammonium sulfate). At the end of 42-h optimal cultivation period, the highest (38.5 g/L) and lowest (28.8 g/L) concentrations of l-lactic acid were obtained with CFP and ammonium sulfate, respectively. This is the first report on use of waste chicken feather as a lactic acid production substrate. In addition, a new R. oryzae strain, being capable of using molasses sucrose as carbon source in order to produce l-lactic acid, was isolated.     

不同碳氮组合对小球藻异养培养油脂积累的影响

目的探讨不同碳氮组合对小球藻Chlorella pro tothecoides CS-41异养培养油脂积累的影响。方法以不同浓度葡萄糖为碳源,尿素为氮源,异养培养小球藻,采用干重法测定小球藻生物量,氯仿/甲醇法提取总脂,并采用GC-MS分析脂肪酸含量。结果小球藻最佳生长条件的碳氮组合配比为葡萄糖浓度40g/L,尿素浓度3g/L,此时小球藻生物量达最高,为9.1g/L,脂肪酸产率最大,为1.26g/L;总脂含量最高,为4.38g/L。结论获得了碳氮最佳配比,使小球藻脂肪酸含量提高到1.26g/L,为发酵扩大化培养小球藻制备生物柴油奠定了基础。     

重组真菌腈水解酶的发酵工艺条件及生物催化特性初探【需加急】

对产真菌腈水解酶的重组大肠杆菌的培养基种类、培养基成分、诱导剂种类和浓度、诱导条件、p H和温度进行了系统考察。摇瓶发酵优化结果显示:以甘油作为主要碳源,蛋白胨和酵母膏作为主要氮源,并添加微量元素的SOC培养基作为发酵培养基,最适接种量为0.5%;较优的诱导剂诱导条件为:采用0.5 mmol/L的IPTG诱导12 h,发酵p H=7.5,诱导温度25℃时产酶效果最佳。经过优化后,重组酶的酶活得到了显著提高,总酶活最高达到了3.84 U/m L,相比初始水平(0.84 U/m L)提高约4倍。5 L发酵罐的放大实验表明,产酶效果良好,总酶活和比酶活均与摇瓶水平基本持平。全细胞催化性质考察研究结果表明,该菌株所产腈水解酶催化反应的最适催化反应温度是45℃,最适反应p H约为7.2。     

Effect of glucose:xylose ratio on xylose reductase and xylitol dehydrogenase activities from Candida guilliermondii in sugarcane bagasse hydrolysate

The influence of glucose on xylose reductase (XR) and xylitol dehydrogenase (XDH) enzyme activity was evaluated from sugarcane bagasse hydrolysate fermentations with different glucose:xylose ratios (1:25, 1:12, 1:5 and 1:2.5) by employing an inoculum of Candida guilliermondii grown in media containing glucose, a mixture of glucose and xylose, or only xylose as carbon sources. According to the results, the glucose:xylose ratio affected positively this bioconversion and a correlation was not observed between the favourable conditions for xylitol production and the XR and XDH activities. Also, the results were influenced not only by the glucose:xylose ratio in the fermentation medium, but also by the carbon source employed in the growth medium of the inoculum. The optimum condition for xylitol production by C. guilliermondii in sugarcane bagasse hemicellulosic hydrolysate should use hydrolysate with a 1:5 glucose:xylose ratio and inoculum grown in medium containing xylose as the only carbon source. Copyright © 2006 Society of Chemical Industry     

The effects of culture condition on the growth property and docosahexaenoic acid production from Thraustochytrium aureum ATCC 34304

The effect of the composition of artificial sea water (ASW) medium on the growth properties and docosahexaenoic acid (DHA) production from Thraustochytrium aureum ATCC 34304 was investigated. A maximum dry cell weight (DCW) of 3.71 g/l was obtained when the NaCl and MgSO₄ concentration in the ASW media were 15 and 0 g/l, respectively. The maximum initial specific growth (ISG) rate of 0.16 was attained at 7.5 g/l NaCl and 18 g/l MgSO₄, while the minimum ISG rate (0.02) was obtained at 5.0 g/l NaCl and 4.5 g/l MgSO₄. The least doubling time required for biomass production was 4.3 h at 7.5 g/l NaCl and different MgSO₄ concentrations. A maximum of 7.9 g/l DCW was obtained on the fourth day of cultivation at 30 g/l glucose and 2.5 g/l (each) yeast extract (YE) and peptone. The DHA content in the lipids was significantly affected by the concentration of glucose and nitrogen sources (YE and peptone) in the ASW medium. At the lowest glucose (10 g/l) and YE/peptone (0.5 g/l) concentration and highest glucose (30 g/l) and YE/peptone (2.5 g/l) concentration, the DHA content was 34.725 and 40.33%, respectively, relative to total lipid content. However, the DHA content in the lipid was not affected by the NaCl and MgSO₄ concentration. At the lowest NaCl (2.5 g/l) and MgSO₄ (4.5 g/l) concentration and highest NaCl (60 g/l) and MgSO₄ (18 g/l) concentration the DHA content was 39.62 and 38.48%, respectively. The maximum DHA content in the lipid was 49.01% after four days of cultivation when 7.5 g/l NaCl and 4.5 g/l MgSO₄ were in the ASW medium. The growth properties of T. aureum ATCC 34304 for biomass production and DHA yield in the lipid content were found to be affected by NaCl and glucose concentration.     

硫磺菌菌丝液体发酵适宜培养基配方研究

实验以硫磺菌为材料,在适宜的外部培养条件（培养温度为25℃、装液量75 mL/250 mL、摇瓶转速为150 r/min培养120 h）下,通过选取不同氮源（包括黄豆浆、尿素、牛肉膏、蛋白胨、（NH4）2SO4）、不同碳源（葡萄糖、蔗糖、淀粉、乳糖）、酸碱度几个单因素的控制以及正交实验,采用液体发酵技术培养菌丝体,以菌丝体湿重为指标,确定了硫磺菌的液体培养的适宜培养基配方。结果表明：以17.5 g/L的黄豆浆为氮源,20 g/L的葡萄糖为碳源,初始pH为5.5的配方既能获得高产又较为方便经济。     

Characterization of Novel Triacylglycerol Estolides from the Seed Oil of Mallotus philippensis and Trewia nudiflora

Triacylglycerol estolides have been reported as components of the seed oil of a number of plant species and are generally associated with the presence of fatty acids containing hydroxyl groups. We have used MALDI-TOF MS to examine the intact acylglycerol species present in the seed oils of two plants that produce kamlolenic acid (18-hydroxy-Δ9cis,11trans,13trans-octadecatrienoic acid). Mallotus philippensis and Trewia nudiflora were both shown to produce seed oil rich in TAG-estolides. Analysis by MALDI-TOF MS/MS demonstrated that the TAG-estolides had a structure different to that previously proposed after enzymatic digestion of the oil. Acylglycerols containing up to 14 fatty acids were detected but fatty acid estolides were only present in a single position on the glycerol backbone, with predominantly non-hydroxyl fatty acids in the remaining two positions. Increased numbers of fatty acids per glycerol backbone were accounted for by the presence of fatty acid estolides containing a correspondingly greater number of fatty acids. For example, acylglycerols containing seven fatty acids had a fatty acid estolide of five fatty acids at one position on the glycerol backbone. Both capped and uncapped fatty acid estolides, with a free hydroxyl group, were present, with capped fatty acid estolides being more abundant in T. nudiflora and uncapped fatty acid estolides in M. philippensis.     

Chiral resolution of differently substituted racemic acetyl‐1‐phenyl ethanol using lipase from Bacillus subtilis

In the present investigation a Lipase producing strain, Bacillus subtilis (MTCC‐121) was grown on various media containing different sources of carbon, nitrogen and other nutrients. The best media found for the production of lipase was M2 media containing 0.4% peptone, 0.2% beef extract and 1% NaCl. Lipase produced from this culture was used for the kinetic resolution of racemic acetyl‐1‐phenyl ethanol and its derivatives, which are important as chiral auxiliaries and intermediates in the synthesis of natural products, pharmaceuticals and agrochemicals. The lipase resolved these substrates after 48 h with enantiomeric excess of 90–98% and conversion 40–48%. Copyright © 2010 Society of Chemical Industry     

Evaluation of a novel low-cost culture medium containing exclusively milk,potato and glycerol for microbial transglutaminase production by Streptomyces mobaraensis

The aim of this work was to evaluate the production of microbial transglutaminase (MTG) by Streptomyces mobaraensis in a reduced nutritional medium based on non-commercial potatoes. Initially, the production was studied comparing 3 culture media based on potato: (i) media based on acid hydrolysed potato, (ii) media based on gelified non-hydrolysed potato and (iii) media based on gelified non-hydrolysed potato supplemented with glucose (20 g/L). All media were supplemented with nutrients and inductors of MTG production (yeast extract, peptone, sodium caseinate, glycerol and mineral salts such as sodium phosphate, potassium phosphate, magnesium sulphate). The best medium was gelified non-hydrolysed potato allowed to obtain up to 2.72 U/mL at 96 h. In a second study, it was proposed the substitution of all the nutrients and inductors by exclusively skim milk and glycerol. An experimental design was performed to optimize the composition of milk, potato and glycerol. The economical yield of the process was also evaluated. Fermentations were carried out and samples were taken at 72 h and 96 h. Results showed that the maximum MTG activity was obtained at 72 h. A significant effect of potato and milk on MTG activity was observed, including an effect of interaction between both compounds. Optimal conditions selected were: 600 g/L of skim milk, 40 g/L of potato and 5 g/L of glycerol, yielding a maximum activity of 3.2 U/mL and an economic yield of 8.11 € of MTG/€ of nutrients. The results showed that the medium milk–potato–glycerol was feasible for the production of transglutaminase, obtaining a high MTG activity in a simple natural medium.     

Urea adducts of mono- and diesters of fatty acids

Summary The experimental work has shown how molecular configurations of fatty acid glycerides affect urea complex formation. Extraction data indicate that urea will form a complex with glycerol monopalmitate, monostearate, and mono-oleate but not under these conditions with glycerol monolinoleate or monolinolenate from mixtures containing these monoesters. Data also indicate that it is easier for urea to form a complex with monoglycerides than with diglycerides. In a mixture containing mono- and diglycerides of saturated and unsaturated fatty acids, urea will separate first on the basis of saturation, then secondly on the basis of the degree of esterification of the glycerol. An erratum to this article is available at .     

Production of Polyunsaturated Fatty Acids by Mortierella alpina Using Submerse and Solid State Fermentation

A new isolate of Mortierella alpina, > 98 % identical with M. alpina ATCC 16266, was cultivated in a defined glucose‐based medium with three organic nitrogen sources (glycine, urea and Na‐L‐glutamate) at three different concentrations in shaking flasks at 20 °C. The results were compared to the cultivation in complex medium with yeast extract as nitrogen source. In the defined media, high yields of polyunsaturated fatty acids (PUFAs) and arachidonic acid (ARA), respectively, were obtained with Na‐L‐glutamate. However, the absolute highest yields of PUFA and ARA were measured with the yeast extract medium. An optimized yeast extract complex medium was used for a submerse bioreactor cultivation in a 45‐L scale. Furthermore, M. alpina was cultivated in a solid state fermenter, using an oat bran water mixture as substrate.     

Effect of subacute toxic levels of dietary cyclopropenoid fatty acids upon membrane function and fatty acid composition in the rat

The effect of subacute toxicity levels of dietary cyclopropenoid fatty acids upon several physiological parameters was determined in the rat. Diets containing 2% corn oil, 2%Sterculia foetida oil or 2% hydrogenatedSterculia foetida oil were fed.Sterculia foetida oil (50% cyclopropenoid fatty acids) fed rats exhibited retarded growth, elevated organ to body wt ratios, increased saturation of tissue lipid, and abnormal histopathology when compared to corn oil and hydrogenatedSterculia foetida oil fed rats. Growth was retarded 50%, liver/body wt doubled, and the percentage of saturated fatty acids in adipose tissue increased 2.5-fold forSterculia foetida oil vs. corn oil comparisons. Three membrane systems were examined in corn oil andSterculia foetida oil fed rats. Erythrocyte hemolysis rate in 0.3 M glycerol was increased by 30%; induction of mitochondrial swelling by reduced glutathione was inhibited completely and microsomal codeine demethylase activity was depressed nearly 50% inSterculia foetida oil fed rats. The ability of cyclopropenoid fatty acids to inhibit fatty acyl desaturase and influence tissue and membrane lipid composition is discussed. Most of the detrimental effects observed in cyclopropenoid fatty acids fed rats may be associated with alteration of normal lipid metabolism and membrane function.     

EFFECT OF NITROGEN AND CARBON SOURCES ON THE INORGANIC PHOSPHATE SOLUBILIZATION BY DIFFERENT ASPERGILLUS NIGER STRAINS

The Mineral phosphate solubilization (MPS) was studied in ten Aspergillus niger strains. MPS activity was measured in solid (Pikovskaya's medium) as well as liquid media using different phosphate sources (tricalcium, dicalcium, ferric and aluminium phosphates, and bone meal), carbon sources (glucose, mannitol, fructose, sucrose, xylose, and sorbitol), and nitrogen sources (ammonium sulfate, ammonium chloride, ammonium nitrate, sodium nitrate, potassium nitrate, and urea). All the strains showed a zone of clearance of tricalcium phosphate in Pikovskaya's medium in plates and solubilized dicalcium and tricalcium phosphates in broth efficiently. Solubilization was lower in ferric and aluminium phosphates. The activity was very poor in bone meal medium. Among the carbon sources the fungi preferred mannitol for higher P solubilization. Nitrogen in the form of nitrate was very effective in solubilizing inorganic phosphates. Xylose and urea were the poorest sources of carbon and nitrogen for all the strains of Aspergillus. Phosphate release was associated with reduction in pH.     

Isolation and Characterization of a Docosahexaenoic Acid-Phospholipids Producing Microorganism Crypthecodinium sp. D31

Thirty-four strains of docosahexaenoic acid (DHA)-producing microorganisms were newly isolated from brackish areas in Japan. These strains showing various compositions of fatty acids. Especially, the fatty acids produced by one of the strains, named D31, had a high DHA content (over 60 % of the total fatty acids) and the simple fatty acid composition (16:0, 18:0, 18:1 and DHA without any other polyunsaturated acids). Although most oleaginous microorganisms accumulate DHA as triacylglycerol, the strain D31 accumulated DHA mainly as a polar lipid (79.4 % of total DHA), especially as phosphatidylcholine (71.4 % of polar DHA). This strain D31 was identified as a related species of Crypthecodinium cohnii on the basis of phylogenetic analysis. Crypthecodinium sp. D31 showed high DHA productivity when cultivated in a medium containing glycerol as the carbon source and a mixture of yeast extract and polypeptone as the nitrogen sources, with a salinity that was equivalent to 50 % of that of seawater and a pH in the acidic range (<pH 6.0). Crypthecodinium sp. D31 is considered as a promising producer of high-purity DHA-containing phospholipids.     

Production of eicosapentaenoic acid byMortierella fungi

Mycelia of arachidonic acid-producing fungi belonging to the genusMortierella were found to be rich sources of 5,8,11,14,17-cis-eicosapentaenoic acid (EPA). Production of EPA by these fungi was observed only when they were grown at low temperature (6–16 C). EPA comprised 5–20% of the total extractable mycelial fatty acids in most strains tested. No significant accumulation of EPA was observed on incubation at high temperature (20–28 C), at which the other major mycelial C-20 fatty acid, arachidonic acid, was still efficiently produced. In a study on the optimization of the culture conditions for EPA production by a selected fungiM. alpina 20–17, a medium containing glucose and yeast extract as major carbon and nitrogen sources, respectively, was found to be suitable. Periodic feeding of glucose during growth of the fungus and cultivation at high temperature (20 C) during the early growth phase followed by temperature shift to 12 C were found to be effective at increasing mycelial yield and reducing cultural period, respectively. Under the optimal culture conditions, the EPA production reached 0.49 mg/ml of culture broth (29 mg/g dry mycelia). This value accounted for 13.5% of the total fatty acids in the extracted lipids. Other major fatty acids in the lipids were palmitic acid (6.0%, by weight), stearic acid (5.3), oleic acid (6.2), linoleic acid (3.0), γ-linolenic acid (3.5) and arachidonic acid (60.0). On leave from Suntory Ltd.     

EFFECT OF NITROGEN AND CARBON SOURCES ON THE INORGANIC PHOSPHATE SOLUBILIZATION BY DIFFERENT ASPERGILLUS NIGER STRAINS

The Mineral phosphate solubilization (MPS) was studied in ten Aspergillus niger strains. MPS activity was measured in solid (Pikovskaya's medium) as well as liquid media using different phosphate sources (tricalcium, dicalcium, ferric and aluminium phosphates, and bone meal), carbon sources (glucose, mannitol, fructose, sucrose, xylose, and sorbitol), and nitrogen sources (ammonium sulfate, ammonium chloride, ammonium nitrate, sodium nitrate, potassium nitrate, and urea). All the strains showed a zone of clearance of tricalcium phosphate in Pikovskaya's medium in plates and solubilized dicalcium and tricalcium phosphates in broth efficiently. Solubilization was lower in ferric and aluminium phosphates. The activity was very poor in bone meal medium. Among the carbon sources the fungi preferred mannitol for higher P solubilization. Nitrogen in the form of nitrate was very effective in solubilizing inorganic phosphates. Xylose and urea were the poorest sources of carbon and nitrogen for all the strains of Aspergillus. Phosphate release was associated with reduction in pH.     

不同C源对丙酮丁醇梭菌产丁醇的影响

对丙酮丁醇梭菌在以葡萄糖、木糖、蔗糖、混合糖、玉米芯酸解糖液分别作C源的P2培养基中的产丁醇状况进行研究。结果表明:不同C源对丙酮丁醇梭菌发酵产丁醇有显著的影响;葡萄糖为底物时,丁醇产量最高达到13.50 g/L,总溶剂为19.66 g/L;蔗糖为底物时,丁醇所占比例都在70%以上,丁醇产量可达12 g/L;木糖、混合糖为底物时,丁醇产量在10 g/L左右;只有丙酮丁醇梭菌I4-28能利用玉米芯酸解糖液发酵产丁醇,丁醇产量为7 g/L。     

Fermentative production of puromycin by Streptomyces alboniger

Two strains of Streptomyces alboniger NRRL B-1832 and 2403 were used for the biosynthesis and production of puromycin using different media. Streptomyces alboniger NRRL B-2403 produced more puromycin than NRRL B-1832 on medium composed of (amounts in g/1), soybean meal 10.0; corn steep 20.0; dextrin 10.0; NaCl 5.0; CaCO₃ 2.0, and K₂HPO₄ 2.0. When the carbon and nitrogen sources of the basal medium were replaced by different carbon and nitrogen sources it was found that dextrin, sucrose, starch and maltose favoured the production of higher titres of puromycin, while generally organic nitrogen sources such as soybean meal and fodder yeast (40.0 and 50.0% total nitrogen) stimulated puromycin yield. On the evidence of their chemical compositions and because they are cheap and produced locally Egyptian black, strap molasses and fodder yeast are preferably used in medium for the production of puromycin. The other conditions favouring and controlling the production and biosynthesis of puromycin by fermentation were investigated.