Amylases, nucleic acids encoding them and methods for making and using them

Patent metadata

Field	Value
Publication number	US-9701950-B2
Application number	US-201514719547-A
Country	US
Kind code	B2
Filing date	May 22, 2015
Priority date	Feb 21, 2001
Publication date	Jul 11, 2017
Grant date	Jul 11, 2017

Abstract

In one aspect, the invention is directed to polypeptides having an amylase activity, polynucleotides encoding the polypeptides, and methods for making and using these polynucleotides and polypeptides. In one aspect, the polypeptides of the invention can be used as amylases, for example, alpha amylases, to catalyze the hydrolysis of starch into sugars.

First claim

What is claimed is:

1. A method of generating a variant amylase comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO:2 comprising:
(a) providing parent clones comprising the nucleic acid sequence as set forth in SEQ ID NO:6, SEQ ID NO:66, and SEQ ID NO:68;
(b) providing primers and amplifying the primers to generate pools of fragments of parent clones;
(c) combining and ligating the pools of fragments of (b) to generate the variant amylase genes;
(d) transform the variant amylase genes of (c) into a host cell that expresses the variant amylase.

2. The method of claim 1 , wherein the primers comprise any one of the nucleic acid sequences of: SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, and SEQ ID NO:65.

Assignees

• Basf Enzymes Llc

Inventors

• Callen Walter
• Richardson Toby
• Frey Gerhard
• Miller Carl A
• Kazaoka Martin
• Mathur Eric J
• Short Jay M

Classifications

• C12P19/14

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• C12N9/2411 Primary

  Amylases · CPC title

• C12N9/2417

  from microbiological source · CPC title