Liquid detergent compositions containing alpha-amylase
A technology of liquid detergent and composition, which is applied in the direction of detergent composition, non-surface active detergent composition, detergent compounding agent, etc., and can solve problems such as bad odor
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[0145] Preparation of the alpha-amylase of the present invention
[0146] The alpha-amylase (variant) of the detergent composition can be prepared using any mutagenesis procedure known in the art, eg, site-directed mutagenesis, synthetic gene construction, semi-synthetic gene construction, random mutagenesis, shuffling, and the like. Site-directed mutagenesis is a technique of introducing one or more (eg, several) mutations at one or more defined sites in a polynucleotide encoding a parent.
[0147]Site-directed mutagenesis can be accomplished in vitro by PCR involving the use of oligonucleotide primers containing the desired mutation. In vitro site-directed mutagenesis can also be performed by cassette mutagenesis, which involves cleavage by restriction enzymes at sites in the plasmid containing the polynucleotide encoding the parent and subsequent oligonucleotides containing the mutation. linked in a polynucleotide. Typically, the restriction enzymes that digest the plas...
Embodiment 1
[0212] Example 1 - Determination of pH profile.
[0213] When using SEQ ID NO: 1 for numbering, the pH profile was determined for the alpha-amylase of SEQ NO: 1 with the substitutions G46A+T47I+G105A+I199F.
[0214] Reducing sugar determination:
[0215] Assay for measuring amylolytic activity (alpha-amylase activity).
[0216] Alpha-amylase activity can be determined by a method using a starch substrate (eg, rice starch S7260 from Sigma-Aldrich). Alpha-amylase cleaves the substrate and the reducing end formed by the amylase is determined by reaction with p-hydroxybenzoic acid hydrazide (reducing sugar assay). This leads to the formation of a hydrazone, which can be detected by absorbance measurement at 405 nm. The number of reducing ends formed is proportional to the amylase activity.
[0217] Enzymes were diluted in enzyme dilution buffer (10 mM MES, pH 7.0, 0.01% TritonX100, 0.100 mM CaCl2) to enzyme concentrations that gave linear dose-response curves in the assay. pH...
Embodiment 2
[0236] Example 2: Amylase storage stability in liquid detergents.
[0237]A series of liquid detergent combinations were formulated using prior art amylases (Stainzyme and Termamyl Ultra) and amylases with substitutions G46A+T47I+G105A+I199F of SEQ ID NO: 1 (ie amylase SEQ ID NO: 2) thing. The compositions used are given in Table 1. Detergent enzyme samples were placed in glass vials, and the glass vials were then closed. Detergent enzyme samples were incubated at 37°C or 45°C for 2 and 4 weeks, and the same set of samples were frozen to represent fresh samples. After storage, residual amylase activity was then measured using the PNP-G7 amylase activity assay. Amylases tested were Termamyl Ultra, Stainzyme (Novozymes A / S; Krogshoejvej 36; 2880 Bagsvaerd; Denmark) and the amylase of SEQ ID NO:2.
[0238] Table 1 - Liquid Detergent Composition
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[0240] Table 2
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[0242] table 3
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[0244] Table 4
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[0246] Under the conditions ...
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