Lipase variants

A technology of lipase and variants, applied in the field of lipase

Active Publication Date: 2009-02-18
NOVOZYMES AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thus, it is known that laundry detergents containing lipase may som

Method used

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  • Lipase variants
  • Lipase variants
  • Lipase variants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0133] enzyme production

[0134] A plasmid containing the gene encoding lipase is constructed and transformed into a suitable host cell using standard methods in the art.

[0135] Fermentations were carried out as fed-batch fermentations using constant medium at a temperature of 34°C and an initial volume of 1.2 liters. The initial pH of the medium was set to 6.5. Once the pH increased to 7.0, by adding 10% H 3 PO 4 to keep this value. Dissolved oxygen levels in the medium were controlled by varying the rate of agitation and using a fixed aeration rate of 1.0 liters of air per liter of medium per minute. The feed addition rate was maintained at a constant level throughout the fed-batch phase.

[0136] The batch medium contains maltose syrup as carbon source, urea and yeast extract as nitrogen source, and a mixture of trace metals and salts. Feeds added continuously during the fed-batch phase contained maltose syrup as a carbon source, while yeast extract and urea were a...

Embodiment 2

[0139] AMSA - Automated Mechanical Stress Test - Used to calculate Relative Performance (RP)

[0140] Enzyme variants of the present application were tested using the Automatic Mechanical Stress Assay (AMSA). Using the AMSA test, it is possible to examine the wash performance of large quantities of small volume enzyme detergent solutions. The AMSA plate has multiple slots for the test solution, and a cover that compresses the textile sample to be washed against all slot openings. During the wash time, the plate, test solution, textile and cover are shaken vigorously to bring the test solution into contact with the textile and to apply mechanical stress. For further description see WO 02 / 42740, especially the paragraph "Special method embodiments" on pages 23-24. The container containing the detergent test solution consists of a cylindrical hole (diameter 6 mm, depth 10 mm) in a metal plate. The soiled fabric (test material) was placed on top of the metal plate and used as a...

Embodiment 3

[0162] GC - Gas Chromatography - for calculating risk factors

[0163] Butyrate release from lipase washed samples was measured by Solid Phase Micro Extraction Gas Chromatography (SPME-GC) using the following method. Four textile pieces (5 mm in diameter) washed in the solutions specified in Table 3 containing 1 mg / L lipase were transferred to gas chromatography (GC) vials. Samples were analyzed on a Varian 3800 GC equipped with a Stabilwax-DA w / Integra-Guard column (30m, 0.32mm ID and 0.25 micron df) and Carboxen PDMS SPME fiber (75 micron). Each sample was preincubated at 40°C for 10 minutes, followed by 20 minute sampling of the head space above the textile sheet with SPME fibers. The sample was then injected onto the column (injector temperature = 250°C). Column flow rate = 2ml helium / min. Column furnace temperature gradient: 0 minutes = 40°C, 2 minutes = 40°C, 22 minutes = 240°C, 32 minutes = 240°C. Butyric acid was detected by FID detection, and the amount of butyric...

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Abstract

The invention provides variant lipases, preferably, variants with reduced tendency to odor generation obtained by introducing mutations in one or more regions identified in a parent lipase.

Description

[0001] Invention Territory [0002] The present invention relates to lipase variants. Background technique [0003] Lipases are useful, for example, as detergent enzymes for removing lipid or fatty stains from clothing and other textiles, and as dough additives for bread and other bakery products. Therefore, the lipase derived from Thermomyces lanuginosus (synonym Humicolalanuginosa, EP 258 068 and EP 305 216) is known under the trade name Lipolase (a product of NovoNordisk A / S) is sold for detergent use. WO 0060063 describes variants of the Thermomyces pilosa lipase which have particularly good first-wash performance in detergent solutions. WO 9704079, WO 9707202 and WO 0032758 also disclose variants of T. pilosa lipase. [0004] In some applications it is of interest to minimize the formation of odor producing short chain fatty acids. Thus, it is known that laundry detergents containing lipase may sometimes leave a residual odor on fabrics soiled with milk (EP 430315). ...

Claims

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Application Information

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IPC IPC(8): C12N9/20
CPCC12N9/20C12N9/18
Inventor 杰斯珀·文德于尔根·C·F·诺策尔金·博尔奇阿兰·斯文德森托马斯·H·卡利森德比·亚弗马兹·E·比约恩瓦德彼得·K·汉森戈海燕迈克尔·拉姆萨
Owner NOVOZYMES AS
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