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An efficient and improved methodology for the screening of industrially valuable xylano-pectino-cellulolytic microbes.

Singh A, Kaur A, Dua A, Mahajan R - Enzyme Res (2015)

Bottom Line: The probability of getting the desired combination was low, so efforts were made to further improve this cost effective methodology for obtaining the high yield of the microbes capable of producing desired combination of enzymes.By inclusion of multiple enrichment steps in sequence, using only practically low cost substrates and without any nutrient media till primary screening stage, this improved novel protocol for screening gave only the desired microorganisms with xylano-pectino-cellulolytic activity.Using this rapid, efficient, cost effective, and improved methodology, microbes with required combination of enzymes can be obtained and the probability of getting the desired microorganisms is cent percent.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Kurukshetra University, Kurukshetra 136 119, India.

ABSTRACT
Xylano-pectino-cellulolytic enzymes are valuable enzymes of the industrial sector. In our earlier study, we have reported a novel and cost effective methodology for the qualitative screening of cellulase-free xylano-pectinolytic microorganisms by replacing the commercial, highly expensive substrates with agricultural residues, but the microorganisms with xylanolytic, pectinolytic, cellulolytic, xylano-pectinolytic, xylano-cellulolytic, pectino-cellulolytic, and xylano-pectino-cellulolytic potential were obtained. The probability of getting the desired combination was low, so efforts were made to further improve this cost effective methodology for obtaining the high yield of the microbes capable of producing desired combination of enzymes. By inclusion of multiple enrichment steps in sequence, using only practically low cost substrates and without any nutrient media till primary screening stage, this improved novel protocol for screening gave only the desired microorganisms with xylano-pectino-cellulolytic activity. Using this rapid, efficient, cost effective, and improved methodology, microbes with required combination of enzymes can be obtained and the probability of getting the desired microorganisms is cent percent. This is the first report presenting the methodology for the isolation of xylano-pectino-cellulolytic positive microorganisms at low cost and consuming less time.

No MeSH data available.


Related in: MedlinePlus

Zone of substrate hydrolysis shown on (a) wheat bran; (b) xylan; (c) citrus peel; (d) pectin; (e) waste paper; (f) CMC by xylano-pectino-cellulolytic bacterial isolate AVS13 and zone of substrate hydrolysis produced by a known xylanase producer Bacillus stearothermophilus on (g) wheat bran, (h) xylan; a known pectinase producer Bacillus subtilis on (i) citrus peel, (j) pectin; and a known cellulase producer Flavobacterium bolustinum on (k) waste paper, (l) CMC.
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fig1: Zone of substrate hydrolysis shown on (a) wheat bran; (b) xylan; (c) citrus peel; (d) pectin; (e) waste paper; (f) CMC by xylano-pectino-cellulolytic bacterial isolate AVS13 and zone of substrate hydrolysis produced by a known xylanase producer Bacillus stearothermophilus on (g) wheat bran, (h) xylan; a known pectinase producer Bacillus subtilis on (i) citrus peel, (j) pectin; and a known cellulase producer Flavobacterium bolustinum on (k) waste paper, (l) CMC.

Mentions: In qualitative analysis, only 0.5% peptone was added to agar plates containing wheat bran, citrus peel, and waste paper individually. Formation of large clear zone around the colonies indicated that the strains were enzymes producer with good substrate hydrolyzing ability. Zone of substrate hydrolysis obtained after Congo red staining on peptone-wheat bran agar plates indicated the xylanolytic nature of the microorganisms. In order to check the efficiency of this methodology, zones were also analysed on peptone agar plate containing commercial xylan. The clearance zones of substrate hydrolysis shown by isolate on peptone agar plate containing wheat bran/xylan (Figures 1(a) and 1(b)) were nearly similar. The highly expensive commercial xylan has been used by several workers [6, 7, 15–17] for the screening of xylanolytic microorganisms. Isolates which showed clear zone of substrate hydrolysis on wheat bran containing plates also gave the substrate clearance zone onto peptone agar medium containing citrus peel, thus indicating the pectinolytic nature of microorganisms. Similarly, comparable zones of substrate hydrolysis by the isolate on peptone agar plate containing citrus peel (Figure 1(c)) or commercial pectin (Figure 1(d)) were obtained. Many workers have reported the use of pectin for qualitative screening of pectinase producing microorganisms [8, 9, 18–20]. Isolates which showed clear zone on wheat bran and citrus peel containing plates were transferred onto waste paper containing plate. The cellulase producing microorganisms also showed zones of substrate hydrolysis of nearly the same size on both peptone-waste paper agar medium (Figure 1(e)) and peptone-cellulose agar medium (Figure 1(f)). Cellulose degrading microbes have been screened by many workers using cellulose/CM-cellulose [10, 21–23].


An efficient and improved methodology for the screening of industrially valuable xylano-pectino-cellulolytic microbes.

Singh A, Kaur A, Dua A, Mahajan R - Enzyme Res (2015)

Zone of substrate hydrolysis shown on (a) wheat bran; (b) xylan; (c) citrus peel; (d) pectin; (e) waste paper; (f) CMC by xylano-pectino-cellulolytic bacterial isolate AVS13 and zone of substrate hydrolysis produced by a known xylanase producer Bacillus stearothermophilus on (g) wheat bran, (h) xylan; a known pectinase producer Bacillus subtilis on (i) citrus peel, (j) pectin; and a known cellulase producer Flavobacterium bolustinum on (k) waste paper, (l) CMC.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4321668&req=5

fig1: Zone of substrate hydrolysis shown on (a) wheat bran; (b) xylan; (c) citrus peel; (d) pectin; (e) waste paper; (f) CMC by xylano-pectino-cellulolytic bacterial isolate AVS13 and zone of substrate hydrolysis produced by a known xylanase producer Bacillus stearothermophilus on (g) wheat bran, (h) xylan; a known pectinase producer Bacillus subtilis on (i) citrus peel, (j) pectin; and a known cellulase producer Flavobacterium bolustinum on (k) waste paper, (l) CMC.
Mentions: In qualitative analysis, only 0.5% peptone was added to agar plates containing wheat bran, citrus peel, and waste paper individually. Formation of large clear zone around the colonies indicated that the strains were enzymes producer with good substrate hydrolyzing ability. Zone of substrate hydrolysis obtained after Congo red staining on peptone-wheat bran agar plates indicated the xylanolytic nature of the microorganisms. In order to check the efficiency of this methodology, zones were also analysed on peptone agar plate containing commercial xylan. The clearance zones of substrate hydrolysis shown by isolate on peptone agar plate containing wheat bran/xylan (Figures 1(a) and 1(b)) were nearly similar. The highly expensive commercial xylan has been used by several workers [6, 7, 15–17] for the screening of xylanolytic microorganisms. Isolates which showed clear zone of substrate hydrolysis on wheat bran containing plates also gave the substrate clearance zone onto peptone agar medium containing citrus peel, thus indicating the pectinolytic nature of microorganisms. Similarly, comparable zones of substrate hydrolysis by the isolate on peptone agar plate containing citrus peel (Figure 1(c)) or commercial pectin (Figure 1(d)) were obtained. Many workers have reported the use of pectin for qualitative screening of pectinase producing microorganisms [8, 9, 18–20]. Isolates which showed clear zone on wheat bran and citrus peel containing plates were transferred onto waste paper containing plate. The cellulase producing microorganisms also showed zones of substrate hydrolysis of nearly the same size on both peptone-waste paper agar medium (Figure 1(e)) and peptone-cellulose agar medium (Figure 1(f)). Cellulose degrading microbes have been screened by many workers using cellulose/CM-cellulose [10, 21–23].

Bottom Line: The probability of getting the desired combination was low, so efforts were made to further improve this cost effective methodology for obtaining the high yield of the microbes capable of producing desired combination of enzymes.By inclusion of multiple enrichment steps in sequence, using only practically low cost substrates and without any nutrient media till primary screening stage, this improved novel protocol for screening gave only the desired microorganisms with xylano-pectino-cellulolytic activity.Using this rapid, efficient, cost effective, and improved methodology, microbes with required combination of enzymes can be obtained and the probability of getting the desired microorganisms is cent percent.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Kurukshetra University, Kurukshetra 136 119, India.

ABSTRACT
Xylano-pectino-cellulolytic enzymes are valuable enzymes of the industrial sector. In our earlier study, we have reported a novel and cost effective methodology for the qualitative screening of cellulase-free xylano-pectinolytic microorganisms by replacing the commercial, highly expensive substrates with agricultural residues, but the microorganisms with xylanolytic, pectinolytic, cellulolytic, xylano-pectinolytic, xylano-cellulolytic, pectino-cellulolytic, and xylano-pectino-cellulolytic potential were obtained. The probability of getting the desired combination was low, so efforts were made to further improve this cost effective methodology for obtaining the high yield of the microbes capable of producing desired combination of enzymes. By inclusion of multiple enrichment steps in sequence, using only practically low cost substrates and without any nutrient media till primary screening stage, this improved novel protocol for screening gave only the desired microorganisms with xylano-pectino-cellulolytic activity. Using this rapid, efficient, cost effective, and improved methodology, microbes with required combination of enzymes can be obtained and the probability of getting the desired microorganisms is cent percent. This is the first report presenting the methodology for the isolation of xylano-pectino-cellulolytic positive microorganisms at low cost and consuming less time.

No MeSH data available.


Related in: MedlinePlus