Abstract
Aspergillus terreus NCFT4269.10 was implemented in solid-state (SSF) and liquid static surface fermentation (LSSF) for biosynthesis of pectinase. Amongst various substrates, like, mustard oil cake, neem oil cake, groundnut oil cake, black gram peels, green gram peels, chickling vetch peels/grass pea peels wheat bran, pearl millet residues, finger millet waste, broken rice, banana peels (BP), apple pomace (AP) and orange peels, banana peel (Musa paradisiaca L.; Family: Musaceae) was most suitable for pectinase biosynthesis (LSSF: 400 ± 21.45 Uml−1; SSF: 6500 ± 1116.21 Ug−1). Optimization of process parameters using one-variable-at-a-time method revealed that an initial medium pH of 5.0 at 30 °C and 96 h of incubation along with mannitol, urea, ammonium persulfate and isoleucine have positive influence on pectinase production. Further, K+ (1 mM), Riboflavin (10 mg 100 ml−1) and gibberellic acid (0.025 %, w/v) supported in enhanced pectinase production. Banana peels and AP at a ratio of 9:1, moisture content of 90 % with 2 % inoculum size were suitable combinations for production of pectinase. Similarly, 96 h of soaking time with 0.1 M phosphate buffer (pH 6.5) is essential for pectinase recovery. Purification to electrophoretic homogeneity revealed 1.42 fold purification with 8.08 % yield and a molecular weight of 24.6 kDa. Scaling up of various fermentation parameters and supplementing BP as the substrate for pectinase production with better recovery could make it promising for different industrial exploitation.
Highlights
In the industrial arena, pectinase, the catch-all idiom that refers to mixtures of primarily three different enzymatic activities [pectin Esterase (PE), polygalacturonase (PG) and pectin/pectate lyase (PL/PAL)] is produced by a variety of bacteria (Kashyap et al 2001) and fungi (Huang and Mahoney 1999)
In Erlenmeyer flask of 150 ml capacity, sterilized fermentation medium (50 ml) having either mustard oil cake (MoC), neem oil cake (NoC), groundnut oil cake (GnoC), black gram peels (BGP), green gram peels (GGP), chickling vetch peels/grass pea peels (CVP), wheat bran (WB), pearl millet residues (PMR), finger millet waste (FMW), broken rice (BR), banana peels (BP), apple pomace (AP), and orange peels (OP) as substrates was inoculated with 1 9 107 spores ml-1 from 7 days old pre-fermentation culture broth and incubated at 30 ± 1 °C under static condition (Sethi et al 2013b)
A native fungal isolate, Aspergillus terrues NCFT 4269.10 was found to be superior in pectinase production as compared to the previously reported results (Patil et al 2012; Maller et al 2013)
Summary
Pectinase, the catch-all idiom that refers to mixtures of primarily three different enzymatic activities [pectin Esterase (PE), polygalacturonase (PG) and pectin/pectate lyase (PL/PAL)] is produced by a variety of bacteria (Kashyap et al 2001) and fungi (Huang and Mahoney 1999). Pectinolytic enzymes are categorized on the basis of their cleavage of the galacturonan portion of the pectin molecule. They can be distinguished between pectinesterases (PE, E.C 3.1.1.11), which modify pectin esters into low methoxyl pectins or pectic acid and pectin deploymerases, that split the glycosidic linkages between galacturonosyl (methyl ester) residues. Polygalacturonases split glycosidic linkage next to free carboxyl groups by hydrolysis, while pectin and pectate lyases split a-1, 4-glycosidic linkages by transelimination ensuing in galacturonide with a double bond between C4
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