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Amylases have wide biotechnological potentials for applications in various industries. An α-amylase-producing bacterium was isolated from deteriorating yam tubers. Molecular characterization using the 16S rRNA gene sequencing was used to confirm the identity of the bacterium as Bacillus subtilis Y25. The effect of some cultural and nutritional fact...
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... The result is similar with the findings of Mohamed (2004) that revealed 3 isoenzymes after ion exchange chromatography. The amylase had purification fold of 4.0 which can be compared with the purification fold of 3.80 of thermostable alpha amylase from Bacillus subtilis (Aladejana et al., 2020). However lower purification fold of 2.67 of amylase from Geobacillus stearothermophilus was reported by (Snehi et al.,2021). ...
Previous studies have demonstrated that probiotic Lactobacillus plantarum CS was able to generate an appreciable amount of extracellular amylase, hence the need to purify and characterize it. The aim of the study was to purify and characterize crude amylase from probiotic Lactobacillus plantarum CS for its industrial applications Three purification steps including ammonium sulphate precipitation, ion exchange chromatography on carboxymethyl sephadex and gel filtration on Sephadex G-75 were utilized. The homogeneity of the purified enzyme was confirmed using sodium deodocyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The purified amylase was characterized on different parameters including substrates hydrolyses, pH and temperature activity and stability profiles. The general purification elution profile revealed two different peaks of amylase activities with outstanding one having a molecular weight of 59.7kDa. Its purification fold was 4.0 with specific activity of 16.44U/mg protein and enzyme yield of 3%. Temperature optimal activity and stability was at 400C and 7.5 for pH activity and stability. Mangenese (Mn2+) (135.17%), tween 80 (128.30%) and some food condiments garlic, thyme, ginger, and tumeric) significantly (p> 0.05) enhanced amylase activity (≥262.40%). However, selenium (Se4+) and hydrogen peroxide (H2O2) were observed to have greatest inhibiting effect (≥30.9%) on the enzyme. Substrate hydrolysis profiles showed that the amylase hydrolyzed all the test starchy substrates with the highest hydrolytic potential on indigenous sweet potato starch (Km value/ Vmax of 1.33mg/ml/ 7.89ml). The rate of hydrolysis of other test substrates had yam> rice>cassava>corn with km values ≤ 4.0mg/ml and Vmax ≤ 25ml. The obtained results gave an insight that amylase produced from Lactobacillus plantarum CS met with the possessed properties suitable for any industrial application especially in food
... Peptone had maximum amylase production with enzyme activity of 22,668 IU ( Figure 2E) compared to other nitrogen sources. Similarly, Simair et al. [36], Acharya et al. [60], Aladejana et al. [61], and Khushk et al. [62] reported maximum amylase production when peptone was used as a nitrogen source. Contrary to this study, beef extract had the maximum production for Bacillus sp. ...
The digestive process and intestinal protein absorption are influenced by a variety of factors. Due to their numerous health advantages, including potential favorable effects on protein digestion and absorption, probiotics have gained increased attention in recent years. Probiotics can control the intestinal microflora, which in turn affects the intestinal bacteria responsible for proteolysis. Additionally, certain probiotics can release exoenzymes that aid in the digestion of proteins and others can stimulate the host’s digestive protease and peptidase activity. By boosting transport and enhancing the epithelium’s capacity for absorption, probiotics can also improve the absorption of tiny peptides and amino acids as well as lessen detrimental protein fermentation, which lowers the toxicity of metabolites. The present study explored the production of enzymes by Bacillus paranthracis strain MHDS3, a probiotic candidate isolated from Pellaea calomelanos. Bacillus paranthracis displayed enzyme activities of amylase (31,788.59 IU), cellulase (4487.486 IU), and pectinase (13.98986 IU) through submerged fermentation. The CAZyme analysis of B. paranthracis revealed 16 CAZyme gene clusters associated with cellulose, amylase, and pectinase activity. Thus, B. paranthracis is a promising probiotic strain that can produce enzymes with biotechnological applications.
... The Vmax and Km obtained were 4834.64 ± 95.71 and 91.09 ± 3.72 Unit/mg protein, respectively. Low Km values for αamylases produced by some Bacillus spp had been reported 24 and indicates that the high affinity of the enzyme with its substrate in the formation of the enzyme-substrate complex. The low Km value obtained from this study has shown robust potentials for harnessing the α-amylase produced by B. cereus SM22 in active biotechnological processes for efficient bioconversions of starch molecules into useful products. ...
Amylases can be sourced through various organisms that thrive on common food items such ascocoyam. This study optimized, purified, and characterized α-amylase from Bacillus cereusSM22 isolated from decayed cocoyam tubers. Healthy cocoyam tubers were homogenized,filtered, allowed to stand, and then decanted. The residue was dried at 50oC for 48 hours,powdered and used as cocoyam starch. Inoculum size, temperature, pH, starch concentration,nitrogen and carbon sources were used to optimize the α-amylase activity/production by Bacilluscereus SM22. The α-amylase activity and protein concentration of the cell-free culturesupernatant were determined. The supernatant was purified on DEAE-Sephacel and BioGel P-100 and the native molecular weight of the α-amylase was estimated. Also, the α-amylase wasoptimized and kinetic parameters; Michaelis constant (Km) and maximum velocity of thereaction (Vmax) were determined. The result showed that pH 7.0, 37oC, 1% cocoyam starch, 2%inoculum, peptone and ammonium nitrate were necessary for α-amylase production by Bacilluscereus SM22. Similarly, pH 6.0, 50oC, NaCl (100 or 240 mM), Mg2+ (80-120 mM) and Ca2+(20-80 mM) were required for maximum α-amylase activity. However, Al3+ and EDTAcompletely inhibited the α-amylase activity. The study successfully purified and optimized α-amylase from Bacillus cereus SM22 isolated from decayed cocoyam tubers. It demonstrated thenecessary factors for optimal performances of Bacillus cereus SM22 and α-amylase in a givenculture medium. Also, the low Km and high Vmax for starch substrate make the α-amylase fromBacillus cereus SM22 an excellent, cheap, and robust candidate for industrial applications
... [3,5]. Alpha-amyase từ các chủng vi khuẩn thuộc loài B. subtilis thường hoạt động tối ưu trong dải pH từ 6 -8 và khoảng nhiệt độ từ 55 đến 60 o C [6][7][8]. ...
... Trong một số trường hợp đặc biệt, một số chủng vi khuẩn B. sublitis có thể sinh trưởng và sinh tổng hợp amylase ở điều kiện nhiệt độ cao: ví dụ như chủng B. sublitis T41a sinh tổng hợp amylase tốt nhất ở nhiệt độ 58 o C [13], chủng B. sublitis Y25 sinh tổng hợp amylase cực đại ở 45 o C [6]. Tuy nhiên, trong đa số các trường hợp thì chủng vi khuẩn thuộc loài B. sublitis thường sinh enzyme tốt nhất ở khoảng nhiệt độ 35 -37 o C [7,12]. ...
... Các kết quả nghiên cứu cho thấy α-amylase từ chủng vi khuẩn B. sublitis V37 bền và hoạt động tốt nhất trong khoảng pH từ 5,0 đến 6,5. Rất nhiều các nghiên cứu trước đây cũng nhận thấy rằng, α-amylase sinh tổng hợp từ B. sublitis bền và hoạt động tốt ở pH từ 5,0 đến 6,5 [7,14,15]. Alpha-amylase hoạt động tốt trong khoảng pH này thường thích hợp trong các ứng dụng liên quan đến công nghiệp sản xuất ethanol, công nghệ thực phẩm, chăn nuôi và dệt may [3,15]. ...
The effect of physical parameters such as temperature and pH on the production, activity, and stability of α-amylase from Bacillus subtilis V37 was investigated. The results indicated that the optimum culture conditions for enzyme activity were pH 7.0 and 35 oC. The optimum pH and temperature for enzyme activity were 6.0 and 70 oC. The crude enzyme was found to be stable in the pH range of 5.0 to 7.0. The enzyme was stable for 1 h at a temperature from 30 to 80 oC; nearly 100% of enzyme activity remained at temperatures of 30 - 40 oC, and about 34% of original activity remained at a temperature of 80 oC. These features demonstrated that α-amylase from B. subtilis V37 can be applied in many areas such as the food, fermentation, and animal feed industries.
Aspergillus flavus isolated from fermented millet flour produced a crude enzyme, which was purified via ammonium sulphate precipitation and subsequent chromatographic techniques. The biochemical characteristics of the purified amylase were thereafter investigated showing activity in a wide range of pH and temperature, with optimal conditions of pH 6.0 and 50 °C. The enzyme retained even 89% of its activity after 1 h at 50 °C and 2 h at pH 6.0. The purified enzyme was stimulated by Ca2+, Zn2+ and Co2+, while Hg2+ and EDTA caused mild inhibition of α-amylase activity. The kinetic indices (Km and Vmax) and molecular weight of the enzyme were estimated in 1.71 mg mL-1, 2.133 μmol min-1 mL-1 and 45 kDa respectively. The catalytic properties of α-amylase from A. flavus makes it a promising candidate for use in various starch processing industries.
İn vitro Şartlarda Double Haploid Bitki
Üretim Yöntemleri
Temperature Metrology
The current study focuses on optimizing, characterizing, and purifying a thermostable amylase from thermophilic bacteria Geobacillus icigianus (BITSNS038). The amylase was produced optimally at 18–24 h of incubation in the presence of starch and tryptone at 7.5 and 3.0 g/L, respectively (pH 7.0, 70°C, and 150 rpm). Km, Vmax values for starch were 2.17 mg/mL and 4.16 U/mL respectively. The enzyme showed excellent thermal stability at 70°C, retaining 62.5% residual activity after 8 h and a reduced deactivation rate constant (kd) of 0.001. When tested for its efficacy in starch hydrolysis, it showed 34.5% hydrolysis of corn starch slurry and antibiofilm activity, but no antimicrobial activity was noticed. The ultrafiltration led to partial purification of amylase showed 2.67-fold purification with a molecular weight in the range of 45 and 66 kDa. This is probably the first-ever report on a thermostable amylase from G. icigianus and its extensive potential uses.
