An extremely alkaline mannanase from Streptomyces sp. CS428 hydrolyzes galactomannan producing series of mannooligosaccharides

Abstract

An alkaline-thermostable mannanase from Streptomyces sp. CS428 was produced, purified, and biochemically characterized. The extracellular mannanase (Mn428) was purified to homogeneity with 12.4 fold, specific activity of 2406.7 U/mg, and final recovery of 37.6 %. The purified β-mannanase was found to be a monomeric protein with a molecular mass of approximately 35 kDa as analyzed by SDS-PAGE and zymography. The first N-terminal amino acid sequences of mannanase enzyme were HIRNGNHQLPTG. The optimal temperature and pH for enzyme were 60 °C and 12.5, respectively. The mannanase activities were significantly affected by the presence of metal ions, modulators, and detergents. Km and Vmax values of Mn428 were 1.01 ± 3.4 mg/mL and 5029 ± 85 µmol/min mg, respectively when different concentrations (0.6-10 mg/mL) of locust bean gum galactomannan were used as substrate. The substrate specificity of enzyme showed its highest specificity towards galactomannan which was further hydrolyzed to produce mannose, mannobiose, mannotriose, and a series of mannooligosaccharides. Mannooligosaccharides can be further converted to ethanol production, thus the purified β-mannanase isolated from Streptomyces sp. CS428 was found to be attractive for biotechnological applications.

Characterization and high-efficiency secreted expression in Bacillus subtilis of a thermo-alkaline β-mannanase from an alkaliphilic Bacillus clausii strain S10

“…strain JAMB-602 [ 9 ] but lower than those extreme alkaline enzymes from Streptomyces sp. CS428 (pH 12.5) [ 22 ], B. subtilis subsp. inaquosorum CSB31 (pH 12.5) [ 53 ], B. halodurans PPKS-2 (pH 11.0) [ 49 ], Bacillus sp.…”

“…MG-33 (0.20 g l −1 ) [ 69 ], N. fischeri P1 (0.83 g l −1 ) [ 32 ], A. nidulans XZ3 (0.90 g l −1 ) [ 25 ], Streptomyces sp. CS428 (1.01 g l −1 ) [ 22 ], and A. niger LW-1 (1.10 g l −1 ) [ 29 ], it was also lower than that of most reported β-mannanases such as those from Bacillus sp. HJ14 (2.20 g l −1 ) [ 70 ], Phialophora sp.…”

“…Only a few Gram negative strains such as Pseudomonas [ 16 ], Vibrio [ 17 ], Thermotoga [ 18 ], Rhodothermus [ 19 ], Klebsiella [ 20 ] and Enterococcus [ 21 ] have been reported to produce β-mannanase. Besides these, some β-mannanases from actinomycetes such as Streptomyces [ 22 ], Cellulomonas [ 23 ] and Thermobifida [ 24 ] have also been studied. For fungi, the most common mannolytic group belongs to the genus Aspergillus [ 25 – 29 ], while some species from genus Penicillium [ 30 ], Trichoderma [ 31 ] and Neosartorya [ 32 ] have also been investigated.…”

Isolasi Bakteri Pendegradasi Xilan dan Manan dari Perairan Indonesia

“…Tidak banyak penelitian yang melaporkan enzim mananase alkali dari bakteri laut, terutama dari perairan Indonesia. Mannanase alkali banyak dilaporkan diproduksi dari Bacillus (Hatada et al, 2005) dan Streptomyces (Yoo et al, 2015;Pradeep et al, 2016) (2014) mengisolasi Bacillus manolitik dari air laut di Pul au Pari, DKI Jakarta untuk produksi manooligosakarida. Potensi mananase alkali sangat luas, seperti dimanfaatkan oleh industri deterjen dan kertas karena sebagian besar proses di industri tersebut dilakukan di pH tinggi (Chauhan, Puri, Sharma, & Gupta, 2012;Dhawan & Kaur, 2007;Hatada et al, 2005).…”

Directed evolution of a β-mannanase from Rhizomucor miehei to improve catalytic activity in acidic and thermophilic conditions

“…So far, numerous β-mannanases from bacteria [8, 9], fungi [10, 11], and actinomyces [12, 13] have been discovered and characterized. Most of them belong to GH families 5 and 26 [14], while only a few β-mannanases are divided into GH families 113 and 134 [9, 11, 12].…”