Extracellularα-Galactosidase fromTrichodermasp. (WF-3): Optimization of Enzyme Production and Biochemical Characterization

Abstract

Trichoderma spp. have been reported earlier for their excellent capacity of secreting extracellular α-galactosidase. This communication focuses on the optimization of culture conditions for optimal production of enzyme and its characterization. The evaluation of the effects of different enzyme assay parameters such as stability, pH, temperature, substrate concentrations, and incubation time on enzyme activity has been made. The most suitable buffer for enzyme assay was found to be citrate phosphate buffer (50 mM, pH 6.0) for optimal enzyme activity. This enzyme was fairly stable at higher temperature as it exhibited 72% activity at 60°C. The enzyme when incubated at room temperature up to two hours did not show any significant loss in activity. It followed Michaelis-Menten curve and showed direct relationship with varying substrate concentrations. Higher substrate concentration was not inhibitory to enzyme activity. The apparent Michaelis-Menten constant (K m), maximum rate of reaction (V max), K cat, and catalytic efficiency values for this enzyme were calculated from the Lineweaver-Burk double reciprocal plot and were found to be 0.5 mM, 10 mM/s, 1.30 U mg−1, and 2.33 U mg−1 mM−1, respectively. This information would be helpful in understanding the biophysical and biochemical characteristics of extracellular α-galactosidase from other microbial sources.

Enhanced elimination of non‐digestible oligosaccharides from soy milk by immobilized α‐galactosidase: A comparative analysis

“…α‐Galactosidases have applications in the food industry for removal of RFOs from various legumes and beans; in the beet‐sugar industry for removal of raffinose from beet sugar; and in the feed industry for improving the nutritional value of animal feed (Boopathy, Gupta, & Ramudu, ; Moon et al, ; Zhou et al, ). Microbial α‐galactosidases are particularly widely applied in the food industries owing to their biochemical properties and catalytic efficiencies (Chauhan, Kumar, Siddiqi, & Sharma, ; Huang et al, ; Katrolia et al, ; Lee et al, ; Wang et al, ). α‐Galactosidases from various microorganisms have been shown to degrade RFOs from soy milk (Ferreira et al, ; Huang et al, ; Jang et al, ; Katrolia et al, ; Saad & Fawzi, ).…”

Properties and applications of α‐galactosidase in agricultural waste processing and secondary agricultural process industries

“…44,74 ⊍-Galactosidases, both acidic and alkaline, have been characterized from bacteria, fungi, and plants; most bacterial ⊍-galactosidases are alkaline with a pH range between 6 and 7.5 whereas fungal, and yeast enzymes are acidic (pH 3.5-4.0) (Table 5). 4,69 The pH range for acidic ⊍-galactosidase varies significantly, with Phaseolus coccineus enzyme catalyzed at pH 3.0, 44 soybean ⊍-galactosidase catalyzed at pH 5.0. 74 ⊍-Galactosidase isolated from bacteria Bacillus sp.…”

“…The availability of α ‐galactosidases that can catalyze reactions at various ranges of temperature and pH levels increases its adoptability into various agricultural processing businesses. The enzymes exhibit stable activity between 30 °C and 50 °C; a few forms can withstand up to 65 °C 21,69 The optimum temperature for enzyme activity depends on the habitat of the source; thermophilic and mesophilic microorganisms display enzyme activity at elevated temperatures, 4 mesophilic fungi, and yeast α ‐galactosidases are active at 50 °C and α ‐galactosidases from Thermatoga sp. and Archeon sp.…”

Enriching Lactobacilli from Fermented Pulse Dal Flour-Analyzing its Efficacy in Utilizing Carbohydrates and Production of α-galactosidase Enzyme During Pigeon Pea Fermentation

“…Although, it is an advantage that this enzyme is extracellular, as it provides high yield, stability and broad-pH range in activity compared to intracellular enzymes. [54][55][56] Furthermore, this isolate was selected for more detailed research on NDO and α-gal activity as it has not been investigated yet from fermented pulse dal flour source isolate, L. brevis (TIP1).…”