Penghasilan CGTase daripada Bacillus sp. G1 berjaya ditingkatkan dengan menggunakan sistem kultur selanjar mengatasi penghasilan daripada kultur kelompok. Aktiviti CGTase tertinggi yang didapati dalam kultur kelompok ialah 28.1 U/ml. Kajian kultur selanjar difokuskan kepada masa kemasukan medium segar yang berbeza (12, 24 dan 48 jam ), kadar pencairan ditetapkan pada 0.03 per jam. Hasil menunjukkan masa memulakan pam medium segar tidak memberi perubahan yang signifikan terhadap aktiviti CGTase (25.7, 26.3 dan 26.1 U/ml masing-masing) dan produktiviti CGTase (0.77, 0.79 dan 0.78 U/ml/j masing-masing) pada keadaan mantap tetapi produktiviti CGTase (0.77 U/ml/j) akan lebih tinggi berbanding produktiviti kultur kelompok apabila masa di antara larian kultur kelompok diambil kira. Malah peningkatan berpotensi ditingkatkan lagi dengan memulakan pam medium segar lebih awal daripada 12 jam dan juga dengan meningkatkan kadar pencairan.
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.
This study was carried out to determine the optimal parameters for the production of biomass of Trichoderma virens UKMP-1M, a fungus isolated from oil-polluted wastewater. The isolate showed maximum growth at day six after incubation in Mineral Salt Medium (MSM) in the presence of 3% (v/v) heavy Khefji Sour crude oil. Although it grew at pH between 5.0 and 7.0, it grew best at pH 5.5. T. virens UKMP-1M grew at temperatures between 25°C and 35°C, with its highest growth at 30°C. Aeration by agitation at 200 rpm was shown to yield the greatest biomass. Peptone at concentration of 1.5% (w/v) was determined to be a better nitrogen source than urea, potassium nitrate (KNO3), yeast extract, ammonium sulphate ((NH4)2SO4) and ammonium chloride (NH4Cl). Addition of 1% (v/v) crude oil to the MSM medium led to higher biomass production than the addition of 3%, 5%, 7% and 10% (v/v) crude oil. The result also revealed that 40% of total petroleum hydrocarbon (TPH), 100% of pristane and 74% of phytane compounds were degraded after 9 days of incubation at optimal physical and nutrient parameters.
Ciri inokulum bagi pengkulturan kulat oleaginus pencilan tempatan, Cunninghamella bainieri 2A1 dibangunkan dengan mengenal pasti kesan jenis, umur dan saiz inokulum terhadap pertumbuhan, penghasilan lipid dan GLA. Pengkulturan dijalankan pada suhu 30ºC dengan kadar goncangan 250 rpm dalam kelalang goncangan 500 mL yang mengandungi 200 mL medium terhad nitrogen. Inokulum spora didapati lebih sesuai berdasarkan produktiviti penghasilan lipid yang tinggi iaitu 0.71 (g/L/hari) berbanding penggunaan inokulum sel vegetatif vegetatif 24 jam dan 48 jam yang masingmasing memberikan produktiviti hanya 0.51 dan 0.45 (g/L/hari). Selain itu, penghasilan GLA (5.3 × 10-2 g/g biojisim tanpa lipid) dalam kultur yang dimulakan dengan inokulum spora (1 × 105 spora/mL) didapati lebih tinggi sebanyak 23% berbanding inokulum sel vegetatif. Kepekatan spora sebanyak 1 × 103 spora/mL menghasilkan morfologi pellet bersaiz 1.04 mm dan berkadaran dengan kandungan lipid dan GLA masing-masing sebanyak 40% (g/g biojisim) dan 8.34 × 10-2 (g/g biojisim tanpa lipid).
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 Cu2+ 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 30oC 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 Fe3+ 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 Mg2+, 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, Mg2+ and Fe3+ have significant positive effect on the lipid accumulation of this fungus, whereas Mn2+ and Cu2+ exert negative effects in combination.
Pertumbuhan sel secara selanjar pada keadaan mantap telah diperolehi dalam pengkulturan dengan bioreaktor padat. Produktiviti (0.02 g/L/j) bioetanol yang paling tinggi diperolehi adalah pada pengudaraan 0.003 vvm. Produktiviti bioetanol didapati meningkat dengan peningkatan kadar pencairan. Produktiviti tertinggi sebanyak 0.037 g/L/j direkod semasa kadar pencairan (D) 0.05 per jam. Penghasilan bioetanol secara selanjar telah berjaya diselenggarakan pada keadaan tidak 100% anaerobik. Pengudaraan yang terbaik untuk produktiviti bioetanol dalam keadaan seimbang pertumbuhan sel dan penghasilan bioetanol ialah pada 0.003 vvm.
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 GLA 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 ×10-2 (g/g lipid less biomass), respectively
A total of eight strains of Lactobacillus and two strains of Salmonella were isolated from free-range Malaysian chickens intestine. Evaluation based on in vitro studies included aggregation, co-aggregation, growth with bile salts, tolerance to acidic pH, and inhibitory activity were carried out. The isolated Lactobacillus were Lactobacillus fermentum IA, Lactobacillus fermentum IB, Lactobacillus fermentum IC, Lactobacillus fermentum ID, Lactobacillus salivarius subsp. salicinus IE, Lactobacillus salivarius subsp. salicinus IF, Lactobacillus salivarius subsp. salivarius IG, and Lactobacillus spp. IH. The corresponding isolated Salmonella were Salmonella spp. 3B21 and Salmonella spp. 1A12. The ability of aggregation and also tolerance to pH 2.5 are found in Lactobacillus fermentum ID, Lactobacillus salivarius subsp. salicinus IF, Lactobacillus salivarius subsp. salivarius IG, and Lactobacillus spp. IH. The isolate most resistance to 1% bile salts is Lactobacillus fermentum ID but observed to be weak in inhibitory activity against Salmonella spp. The best co-aggregation and strongest inhibitory activity against Salmonella spp. was observed in Lactobacillus salivarius subsp. salivarius IG. Despite being not so resistant in the presence of bile salts 0.5 and 1% (w/v), the lag time in the presence of bile salts 0.3% (w/v) of Lactobacillus salivarius subsp. salivarius IG and also for Lactobacillus spp. IH are the shortest. Based on good aggregation properties, the best co-aggregation, tolerance to acidic pH 2.5 and bile salts 0.3% (w/v) and strongest inhibitory activity against Salmonella spp., Lactobacillus salivarius subsp. salivarius IG comes out as the best candidate as probiotic for chicken.
Delta 6-asid lemak desaturase dan delta 12-asid lemak desaturase merupakan enzim yang diperlukan bagi langkah desaturasi semasa proses biosintesis asid gamma-linolenik (GLA) oleh kulat oleaginus. Objektif kajian ini ialah untuk menganalisis profil pengekspresan gen mengekod enzim delta 6-asid lemak desaturase (des6) dan delta 12-asid lemak desaturase (des12) kulat oleaginus Cunninghamella bainieri semasa penghasilan GLA. Jujukan gen separa bersaiz 1372 pb bagi des6 dan 1008 pb bagi des12 telah dipencil daripada C. bainieri. Analisis pengekspresan gen menggunakan kaedah tindak balas berantai polimerase kuantitatif masa sebenar (RT-qPCR) menunjukkan perubahan kadar pengekspresan des6 adalah lebih tinggi berbanding kadar pengekspresan des12 semasa penghasilan GLA. Pengekspresan des6 adalah tertinggi selepas 24 jam dikultur dalam medium penghasilan GLA. Namun, kadar pengekspresannya menurun hingga jam ke-96 pertumbuhan tetapi meningkat semula pada jam ke-120. Bagi des12, kadar pengekspresannya adalah lebih sekata dengan pengekspresan tertinggi dikesan pada jam ke-120. Analisis penghasilan GLA menunjukkan jumlah GLA dalam sel berkolerasi dengan kadar pengekspresan des6. Hasil kajian mencadangkan bahawa aras pengekspresan des6 adalah penting dalam menentukan aras GLA dalam C. bainieri.
Muhammad Qadri Effendy Mubarak, Siti Hajar Mohamad Jufri, Shikh Mohd Shahrul Nizan Shikh Zahar, Mohd Sahaid Kalil, Aidil Abdul Hamid, Mohd Hafez Mohd Isa
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 (μmax) 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 μmax 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 μmax 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, μmax 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