Cunninghamella bainieri 2A1 is an oleaginous fungus whose lipid accumulation profile is significantly influenced by metal ion concentrations in growth medium. Mg2+, Fe3+, Mn2+ and Cu 2+ were found to be the important elements affecting lipid accumulation in this fungus. This study employs a statistical method (Response Surface Methodology - RSM) to study the combined effects of Mg2+, Fe3+, Mn2+ and Cu2+ on lipid accumulation of C. baineri 2A1. Cultivation was carried out in 250 mL Erlenmeyer flasks containing 100 mL nitrogen limited medium at 30�C and 250 rpm agitation for 120 h. A thirty-run central composite design experiment was employed to identify and optimize the significant factors. In addition to Mg2+ and Fe 3+ which were shown to have significant effects on lipid accumulation, the interactions between Mg2+ and Cu2+, as well as the effect of Cu2+ in quadratic terms were also found to have significant effect on the process (p<0.05). The highest amount of lipid obtained in this study was 39% g/g biomass with optimal levels of Mg 2+, Fe3+ and Cu2+ at 5.00, 0.017 and 0.0005 g/L, respectively, while Mn2+ was omitted. A 32% increment in lipid yield was recorded, where the lipid content increased to 38%, compared to initial yield of 29% g/g biomass prior to optimization. In conclusion, Mg 2+ and Fe3+ have significant positive effect on the lipid accumulation of this fungus, whereas Mn2+ and Cu2+ exert negative effects in combination.

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The influence of the presence of ammonium ions in growth culture on malic enzyme (ME) isoforms activity and lipogenesis in Cunninghamella bainieri 2A1 was investigated. The fungus was cultivated in a nitrogen-limiting medium for 120 h at 30°C under two conditions. One of the cultures was intermittently fed with ammonium tartrate to maintain the ammonium concentrations above 0.5 g/L. The second culture was performed without any feeding to allow N limitation, thus promoting lipid accumulation. Activity staining of ME isoforms was carried out for both cultures. The culture which was not intermittently fed with ammonium tartrate achieved a maximum lipid content of 35% (g/g biomass) at 48 h. This culture possessed five ME isoforms (A, B, C, D and E) with isoform E showing a parallel correlation to lipid accumulation profile. In contrast, intensity of bands representing isoform D decreased as lipid accumulated. No appreciable differences of all other isoforms were observed. However, the culture which was intermittently fed with ammonium tartrate, accumulated only up to 16% lipid (g/g biomass). All isoforms were present but with a more pronounced activity of isoform D and a lower activity of isoform E was observed. These findings support further evidence that isoform E is the key isoform for lipid synthesis in C. bainieri 2A1.

Inoculum for a local isolate of oleaginus fungi, Cunninghamella bainieri 2A1 was developed by establishing the inoculum types, age and size for growth, lipid and gamma linolenic acid (GLA) production. Cultivation was carried out in 500 mL shake flask containing 200 mL of nitrogen limiting medium at 30°C and 250 rpm agitation. Direct inoculation of spores into the production cultures gave higher productivity of lipid production 0.71 (g/L/day) than using 24 and 48 vegetative cells cultures which gave only 0.51 and 0.45 (g/L/day), respectively. Besides, production of GLA (5.3 × 10 -2 g/g lipid less biomass) in the cultures which directly inoculated with spores (1 × 10 5 spores/mL) was 23% higher than those produced in the cultures started with vegetative cells. Spore concentration of 1 × 10 3 spores/mL produced pelletal growth in size of 1.04 mm which contain 40% (g/g biomass) lipid and 8.34 × 10 -2 (g/g lipid less biomass).

A new solvent-producing Clostridium has been isolated from soil used in intensive rice cultivation. The 16S rRNA analysis of the isolate indicates that it is closely related to Clostridium acetobutylicum, with a sequence identity of 96%. The new isolate, named C. acetobutylicum YM1, produces biobutanol from multiple carbon sources, including glucose, fructose, xylose, arabinose, glycerol, lactose, cellobiose, mannitol, maltose, galactose, sucrose and mannose. This isolate can also utilize polysaccharides such as starch and carboxylmethyl cellulose (CMC) for the production of biobutanol. The ability of isolate YM1 to produce biobutanol from agro-industrial wastes was also evaluated for rice bran, de-oiled rice bran, palm oil mill effluent and palm kernel cake. The highest concentration of biobutanol (7.27 g/L) was obtained from the fermentation medium containing 2% (w/v) fructose, with a total acetone-butanol-ethanol (ABE) concentration of 10.23 g/L. The ability of isolate YM1 to produce biobutanol from various carbon sources and agro-wastes indicates the promise of the use of this isolate for the production of biobutanol, a renewable energy resource, from readily available renewable feedstocks. 2015 Elsevier Ltd.

A kinetic model of bacterial growth and metabolite production can adequately explain the trends and interaction of important parameters in a fermentation process. Production of surfactin by two bacterial strains, namely, Bacillus subtilis MSH1 and Bacillus subtilis ATCC 21322, in a 5 L bioreactor was investigated using Cooper's media with 4% (v/v) glucose. The present kinetic study was carried out in order to determine the correlation between microbial cell growth, surfactin production and glucose consumption. Batch fermentation was performed by cultivation of each selected strain in a bioreactor at 30�C for 55 h. The experimental results showed production of surfactin in the culture medium after 5 and 10 h of incubation for B. subtilis ATCC 21332 and B. subtilis MSH1, respectively, at which the bacterial cells were at an early stage of the log phase. The maximum concentration of surfactin (Pmax) achieved by B. subtilis MSH1 and B. subtilis ATCC 21332 was 226.17 and 447.26 mg/L, respectively. The kinetic study of bacterial cell growth of both strains indicated that B. subtilis MSH1 had a specific growth rate (?max) of 0.224 h-1 and attained a maximum biomass concentration (Xmax) as high as 2.90 g/L after 28 h of fermentation, while B. subtilis ATCC 21332, with ?max of 0.087 h-1, attained an Xmax of 2.62 g/L after 45 h of incubation. B. subtilis MSH1 showed higher growth kinetics, thus exhibited higher values of ?max and Xmax compared with B. subtilis ATCC 21332 under identical fermentation conditions. The Pmax achieved by B. subtilis ATCC 21332 was 447.26 mg/L, two times higher than that achieved by B. subtilis MSH1 (226.17 mg/L). The results obtained provide kinetics information including values of Pmax, ?max and Xmax for better understanding of interactions of bacterial cell growth and glucose consumption towards surfactin production by a commercial strain of B. subtilis ATCC 21332 and a local isolate of B. subtilis MSH1.

Thraustochytrids are marine protists belonging to the class Labyrinthula of the kingdom Chromista. This group of microbes is considered a promising alternative source of high-valued omega-3 oils, especially docosahexaenoic acid (DHA, C22:6. n3), replacing fish oil which is the current major source of DHA and eicosapentaenoic acid (EPA, C20:5. n3). Our new isolate, SW1, was considered to be an Aurantiochytrium based on its morphology, fatty acid profile and molecular phylogenetic analysis. Microscopic observations revealed that SW1 has high similarity to Aurantiochytrium limacinum ATCC MYA-1381 (SR21), possessing spherical vegetative cells which undergo repeated bipartition to form diads, tetrads, octads and zoosporangia that release motile zoospores. When cultivated in four different media, SW1 produced the highest biomass (13.17. g/L) and DHA (3.6. g/L) in a medium composed of glucose, sea salt, monosodium glutamate (MSG) and yeast extract. Visualization of lipid droplet development in SW1 using Sudan Black B dye revealed that lipid droplets enlarge to occupy almost the entire cell volume within 72 to 96. h of cultivation. This strain was also found to be able to utilize various saccharides as carbon source. The results of this study show that Aurantiochytrium sp. SW1 is a potential candidate to be developed as commercial microbial DHA producer. � 2015 Elsevier B.V.

The effects of ammonium tartrate and glucose concentration on biomass, lipid and GLA accumulation in Cunninghamella sp. 2A1 were investigated using Response Surface Methodology (RSM). Cultivation was carried out in 250 mL shake flask containing 100 mL of nitrogen limiting medium (with various combinations of concentration of ammonium tartrate (1-3 g/L) and glucose (30-60 g/L) at 30�C and 250 rpm agitation for 120 h. The concentration of both compounds significantly affected the biomass, lipid and CLA yield (p<0.05), with the production of each of them being represented by quadratic models. Higher concentration of ammonium tartrate and glucose (2.99 and 59.33 g/L, respectively) was required for enhanced biomass production whereas low nitrogen content with excess glucose was otherwise favoured for lipid and GLA production. Ammonium tartrate and glucose concentration at 1 and 43 g/L, respectively were estimated by the model and proven to give the highest lipid production and GLA yield of 31.06 % (g/g biomass) and 4.15 x 10-2(g/g lipid less biomass), respectively.

The aim of this study was to establish a simple, accurate and reproducible method for the identification and quantification of surfactin using high-performance liquid chromatography (HPLC). Previously reported method of identification and quantification of surfactin were time consuming and requires a large quantity of mobile phase. The new method was achieved by application of Chromolith® high performance RP-18 (100 × 4.6 mm, 5 μm) as the stationary phase and optimization of mobile phase ratio and flow rate. Mobile phase consisted of acetonitrile (ACN) and 3.8 mM trifluroacetic acid (TFA) solution of 80:20 ratio at flow rate of 2.2 mL/min was obtained as the optimal conditions. Total elution time of the obtained surfactin peaks was four times quicker than various methods previously reported in the literature. The method described here allowed for fine separation of surfactin in standard sample (98% purity) and surfactin in fermentation broth.