# US 11,028,134 B2 · Modified Vip3 polypeptides

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- Number: US 11,028,134 B2
- Document ID: US11028134B2
- Status: Granted
- Kind: B2
- Application number: 16/533,950
- Filed: 2019-08-07
- Published: 2020-01-30
- Granted: 2021-06-08
- Earliest claimed: 2014-08-29
- Assignees: Syngenta Participations AG
- Applicants: Syngenta Participations AG
- Inventors: Joseph M. Watts
- Web page: https://patentgrep.com/patent/US11028134B2

## CPC

- A01N 63/50: Isolated enzymes; Isolated proteins (peptides A01N37/46)
- A01N 37/46: N-acyl derivatives
- A01N 63/10: Animals; Substances produced thereby or obtained therefrom
- C07K 14/195: from bacteria
- C07K 14/32: from Bacillus (G)
- C07K 14/325: Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins
- C12N 15/8279: {for biotic stress resistance, pathogen resistance, disease resistance}
- C12N 15/8286: {for insect resistance}
- C07K 2319/20: containing a tag with affinity for a non-protein ligand
- C07K 2319/55: containing a fusion with a toxin, e.g. diphteria toxin
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The present invention is directed to the vegetative insecticidal proteins (Vips) modified to comprise heterologous bacterial carbohydrate binding modules and the methods of use thereof. Expression of modified proteins resulted in variations to activity against lepidopteran pest species of agricultural importance such as Corn earworm and Fall armyworm conveying broad spectrum insect control.

## Claims

### Claim 1

A modified Vip3A polypeptide comprising a heterologous carbohydrate binding module (CBM), wherein the CBM is from a bacterial β-1,4-mannanase and is substituted for all or a portion of Domain III of a Vip3A polypeptide, and wherein the modified Vip3A polypeptide comprises all or a portion of SEQ ID NO:2 and is pesticidal against an insect.

### Claim 2 (depends on 1)

The modified Vip3A polypeptide of claim 1, wherein the Vip3A polypeptide comprises a portion of SEQ ID NO:2.

### Claim 3 (depends on 1)

The modified Vip3A polypeptide of claim 1, wherein all or a portion of Domain III comprises amino acids 542 to 667 of SEQ ID NO:2.

### Claim 4 (depends on 1)

The modified Vip3A polypeptide of claim 1, wherein the heterologous CBM comprises a metal binding site.

### Claim 5 (depends on 1)

The modified Vip3A polypeptide of claim 1, wherein the modified Vip3A polypeptide is pesticidal against a lepidopteran insect.

### Claim 6 (depends on 1)

The modified Vip3A polypeptide of claim 1, wherein the modified Vip3A polypeptide is pesticidal against at least one of Ostrinia nubilalis (European corn borer), Plutella xylostella (diamondback moth), Spodoptera frugiperda (fall armyworm), Agrotis ipsilon (black cutworm), Agrotis orthogonia (pale western cutworm), Striacosta albicosta (western bean cutworm), Helicoverpa zea (corn earworm), Heliothis virescens (tobacco budworm), Spodoptera exigua (beet armyworm), Helicoverpa punctigera (native budworm), Helicoverpa armigera (cotton bollworm), Manduca sexta (tobacco hornworm), Trichoplusia ni (cabbage looper), Pectinophora gossypiella (pink bollworm), Diatraea grandiosella (southwestern corn borer), Diatraea saccharalis (sugarcane borer), Elasmopalpus lignosellus (lesser cornstalk borer), Psuedoplusia includens (soybean looper), Anticarsia gemmatalis (velvetbean caterpillar), Plathypena scabra (green cloverworm), Homoeosoma electellum (sunflower head moth), and Cochylis hospes (banded sunflower moth), or any combination thereof.

### Claim 7 (depends on 1)

The modified Vip3A polypeptide of claim 1, wherein the heterologous CBM is SEQ ID NO:29.

### Claim 8 (depends on 7)

The modified Vip3A polypeptide of claim 7, wherein the modified Vip3A polypeptide comprises SEQ ID NO:21.

### Claim 9 (depends on 1)

A composition comprising the modified Vip3A polypeptide of claim 1 in an agriculturally acceptable carrier.