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Polypeptides and nucleic acids encoding same

A coding and nucleic acid sequence technology, applied in the field of polypeptides and nucleic acids encoding the polypeptides, can solve problems such as the potential utility of no amino acid sequences

Inactive Publication Date: 2013-07-03
NOVOZYMES AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are no publications of any amino acid sequence that might correspond to this DNA fragment, and thus no indication of any potential utility of the encoded amino acid sequence

Method used

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  • Polypeptides and nucleic acids encoding same
  • Polypeptides and nucleic acids encoding same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 8

[0029] (i) an increase in the number of total facultative anaerobic bacteria in the body, for example in piglets and / or chicks, preferably ileo-rectal contents and / or cecal contents, respectively, in a medium supplemented with 5% vol / vol sheep blood Cultured on Brucella agar, incubated at 37°C in an anaerobic box for 5 days and then measured;

[0030] (ii) an increase in the number of Escherichia coli (Escherichia coli) in vivo, for example in piglets and / or chicks, preferably the ileo-rectal contents and / or cecal contents are respectively treated with Coli-ID pigment producing medium under aerobic conditions measured after incubation at 37°C for 24 hours.

[0031] (iii) The total lactic acid bacteria in the body are substantially unchanged or increased, for example, in piglets and / or chicks, preferably ileo-rectal contents and / or cecal contents, respectively, on MRS agar in an anaerobic box at 37°C Measured after 48 hours of incubation.

[0032] (iv) The total Lactobacillus...

Embodiment 6

[0092] The experiments of Example 6 were PCR reactions, performed in this example with DNA isolated from various strains of Bacillus licheniformis. In a specific embodiment of this assay, the DNA used as template for the PCR reaction is chromosomal DNA that can be isolated by techniques known in the art. The test of Example 6 was positive when a PCR fragment of the correct size was obtained. In Example 6, the correct size is expressed as 0.4kb. In a specific embodiment, the correct size is 0.35kb to 0.44kb (=350bp-440bp). In alternative embodiments, the correct size is 330-430bp, 340-420bp, 350-410bp, 360-400bp, 370-390bp or 385-395bp. The size of the coding sequence (CDS) of SEQ ID NO: 1 is about 380 bp (ie 378 bp).

[0093] nucleic acid sequence

[0094] The present invention also relates to an isolated nucleic acid sequence comprising a nucleic acid sequence encoding a polypeptide (a) encoding a polypeptide of the present invention; (b) hybridizing under very low string...

Embodiment 1

[0221] Example 1: Shake flask fermentation of Bacillus licheniformis

[0222] Bacillus licheniformis ATCC 14580 is propagated overnight at 37° C. on TY agar medium (TY culture fluid solidified with 2% agar), and inoculated into shake flasks comprising 100 ml of TY culture fluid. This TY culture fluid has the following components: Tryptone: 20g / l, yeast extract: 5g / l, FeCl 2 4H 2 O: 7mg / l, MnCl 2 4H 2 O: 1mg / l, MgSO 4 ·7H 2 O: 15 mg / l, pH 7.3. The shake flasks were incubated at 37°C for 20 hours with a shaking speed of 225 rpm. Cells were removed by centrifugation.

[0223] SDS polyacrylamide gel electrophoresis of the supernatant showed a band with a relative molecular weight of 12 kDa consistent with the expected polypeptide. The polypeptide of the present invention is called "L12 protein" because the relative molecular weight of the polypeptide measured by SDS-PAGE is 12 kDa (however, the theoretical molecular weight can be calculated to 9591.56 Da).

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Abstract

The present invention relates to isolated polypeptides and isolated nucleic acid sequences encoding the polypeptides, as well as methods for producing and using the polypeptides in animal feed. An example of a polypeptide of the invention is the so-called L12 protein from Bacillus lichen form is ATCC 14580 which improves the Feed Conversion Ratio (FCR) and / or acts as a gut microflora modulator. The invention furthermore relates to the probiotic use of strains of Bacillus which produce proteins related to L12.

Description

field of invention [0001] The present invention relates to isolated polypeptides as well as isolated nucleic acid sequences encoding said polypeptides. The invention also relates to nucleic acid constructs, vectors and host cells comprising the nucleic acid sequence, as well as methods of producing said polypeptides and using said polypeptides in animal feed, e.g. for improving the Feed Conversion Ratio (FCR) And / or for regulating gut microflora. An example of a polypeptide according to the invention is the so-called L12 protein from Bacillus licheniformis (Bacillus licheniformis) ATCC 14580, which protein has the amino acid sequence of amino acid +1 to +85 of SEQ ID NO: 2 herein (hereinafter amino acid 1 of SEQ ID NO: 2 -85). The present invention also relates to the probiotic use of Bacillus strains producing L12-related proteins in animal feed. Background of the invention Background technique [0002] WO 03 / 093453 discloses in Example 1 the design of an improved Baci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/32C12N9/00C12N15/31C12N15/52A23K1/00A23K10/18A23K20/147
Inventor 彼得·R·奥斯特加德莱恩·N·安德森奥里莉亚·赛翁卡洛斯·西莫斯-努涅斯安娜-玛利亚·克伦特伦纳多·德玛利亚比约恩·E·克里斯坦森
Owner NOVOZYMES AS
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