Detergent compositions containing geobacillus tepidamans mannanase and methods of use thereof
A mannanase, detergent technology, applied in detergent compositions, detergent compounding agents, biochemical equipment and methods, etc., can solve problems such as unsuitability for formulations and washing conditions
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[0252] The following examples are provided to demonstrate and illustrate certain preferred embodiments and aspects of the invention and should not be construed as limiting.
[0253] In the following experimental disclosures, the following abbreviations are used: M (mole / liter), mM (millimol / liter), μM (micromoles / liter), nM (nanomoles / liter), mol (mole), mmol (mmol ), μmol (micromole), nmol (nanomole), g and gm (gram), mg (milligram), μg (microgram), pg (picogram), L (liter), ml and mL (milliliter), μl and μL (microliter), cm (centimeter), mm (millimeter), μm (micrometer), nm (nanometer), U (unit), MW (molecular weight), s (second), min (minute), h (hour ), ℃ (Celsius), QS (sufficient), ND (not performed), rpm (rev / min), H 2 O (water), dH 2 O (deionized water), HCl (hydrochloric acid), aa (amino acid), bp (base pair), kb (kilobase pair), kD (kilodalton), MgCl 2 (magnesium chloride), NaCl (sodium chloride), Ca (calcium), Mg (magnesium), HEPES (4-(2-hydroxyethyl)-1-piperazine...
example 1
[0255] Cloning of Glycosyl Hydrolase Gte Man1 from Geobacillus thermophilicis
[0256] Select Geobacillus thermophile ( et al., IJSEM, 54:2361-2368, 2004 ( et al., International Journal of Systematic and Evolutionary Microbiology, Vol. 54, pp. 2361-2368, 2004)) as a potential source of diverse glycosyl hydrolases and other enzymes for industrial applications. Genomic DNA for sequencing was obtained by incubating Geobacillus thermophilic strain DSM16325 on a heart infusion agar plate (Difco) at 50°C for 24 hours. Cellular material was scraped from the plates and used to prepare genomic DNA with the aid of the ZF Fungal / Bacterial DNA Miniprep Kit from Zymo (Cat# D6005). The genomic DNA was used for genome sequencing and used to amplify the Gte Man1 gene for expression cloning. use Whole genome sequencing of Geobacillus thermophilic strain DSM16325 (obtained from DSMZ, Germany) by sequencing-by-synthesis technology (www.baseclear.com / sequencing / illumina-sequencing / ). G...
example 2
[0323] Expression of Glycosyl Hydrolase Gte Man1 from Geobacillus thermophilicis
[0324] The Gte Man1 gene was amplified from genomic DNA of Geobacillus thermophilicis using the following primers: Primer 1 5'-GGCAGCTGGT AAAAAAAAA CAAAAAAATC CTAGCAAACC-3' (SEQ ID NO: 3), and Primer 2 (XhoI) 5'-CGCCTCGAGT TATTCGGACA ATGCCACGTC AT -3' (SEQ ID NO: 4). The aprE promoter and aprE signal sequence were independently amplified from the p2JM103BBI expression vector using the following primers: Primer 3 5'-TTGTTTTTTT TTACCAGCTG CCTGCGCGCT CA-3' (SEQ ID NO: 5) and Primer 4 (EcoRI) 5'-CGCGAATTCT CCATTTTCTT CTGCTATC -3' (SEQ ID NO: 6) (Vogtentanz, Protein Expr Purif, 55:40-52, 2007 (Vogtentanz, "Protein Expression Purification", Vol. 55, pp. 40-52, 2007)).
[0325] The two PCR products were then assembled into one by a third PCR reaction using primers 2 and 4. After digestion with EcoRI / XhoI, the PCR product was cloned into the p2JM103BBI expression vector digested with the same restri...
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