Polyactive carbohydrates and methods of use thereof
Described herein are biological devices and methods for using the same to produce a polyactive carbohydrate. The biological devices include microbial cells transformed with a DNA construct containing genes for producing a chitin synthase, a chitosanase, 5 and a chitin deacetylase, and, optionally, lipase, regulatory sequence CHR1, transglycosylase, dehydrogenase, and/or (1→3), (1→4)-β-glucan synthase. Methods for using the polyactive carbohydrate are also provided herein including, but not limited to, increasing plant hormone production and improving the appearance and root strength of plants and production of polyurethane biofoams.
1 . A method for increasing root growth of a plant, the method comprising contacting the plant with an extract produced from a biological device, wherein the biological device comprises host cells transformed with a DNA construct comprising the following genetic components: (a) a gene that expresses chitin synthase, (b) a gene that expresses chitosanase, and (c) a gene that expresses chitin deacetylase, wherein the extract comprises a polyactive carbohydrate.
2 . A method for increasing the drought-resistance of a plant, the method comprising contacting the plant with an extract produced from a biological device, wherein the biological device comprises host cells transformed with a DNA construct comprising the following genetic components: (a) a gene that expresses chitin synthase, (b) a gene that expresses chitosanase, and (c) a gene that expresses chitin deacetylase, wherein the extract comprises a polyactive carbohydrate.
3 . A method for improving the physical appearance of a plant, the method comprising contacting the plant with an extract produced from a biological device, wherein the biological device comprises host cells transformed with a DNA construct comprising the following genetic components: (a) a gene that expresses chitin synthase, (b) a gene that expresses chitosanase, and (c) a gene that expresses chitin deacetylase, wherein the extract comprises a polyactive carbohydrate.
4 . The method according to claim 1 , wherein the extract is further applied to soil where the plant is growing.
5 . The method according to claim 1 , wherein the plant is further contacted with chitosan.
6 . The method of claim 5 , wherein the plant is contacted with a composition comprising the extract and chitosan.
7 . The method according to claim 1 , wherein the gene that expresses chitin synthase comprises SEQ ID NO. 2 or at least 70% homology thereof.
8 . The method according to claim 1 , wherein the gene that expresses chitosanase has comprises SEQ ID NO. 3 or at least 70% homology thereof.
9 . The method according to claim 1 , wherein the gene that expresses chitin deacetylase comprises SEQ ID NO. 4 or at least 70% homology thereof.
10 . The method according to claim 1 , wherein the construct further comprises a gene that expresses lipase.
11 . The method of claim 10 , wherein the gene that expresses lipase comprises SEQ ID NO. 1 or at least 70% homology thereof.
12 . The method according to claim 1 , wherein the construct further comprises a gene that expresses (1→3), (1→4)-β-glucan synthase.
13 . The method of claim 12 , wherein the gene that expresses (1→3), (1→4)-β-glucan synthase comprises SEQ ID NO. 6 of at least 70% homology thereof.
14 . The method according to claim 1 , wherein the construct further comprises a gene that expresses chitin synthase regulatory factor CHR1.
15 . The method of claim 14 , wherein the gene that expresses chitin synthase regulatory factor CHR1 comprises SEQ ID NO. 11 or at least 70% homology thereof or SEQ ID NO. 16 or at least 70% homology thereof.
16 . The method according to claim 1 , wherein the construct further comprises a gene that expresses transglycosylase.
17 . The method of claim 16 , wherein the gene that expresses transglycosylase comprises SEQ ID NO. 12 or at least 70% homology thereof or SEQ ID NO. 15 or at least 70% homology thereof.
18 . The method according to claim 1 , wherein the construct further comprises a gene that expresses a dehydrogenase.
19 . The method of claim 18 , wherein the gene that expresses a dehydrogenase expresses a protein comprising SEQ ID NO. 13 or at least 70% homology thereof.
20 . The method according to claim 1 , wherein the construct further comprises one or more ribosomal binding sites preceding the gene that expresses lipase, the gene that expresses chitin synthase, the gene that expresses chitosanase, the gene that expresses chitin deacetylase, the gene that expresses (1→3), (1→4)-β-glucan synthase, the gene that expresses chitin synthase regulatory factor CHR1, the gene that expresses transglycosylase, or any combination thereof.
21 . The method according to claim 1 , wherein the construct further comprises a gene that confers resistance to an antibiotic.
22 . The method according to claim 1 , wherein the construct further comprises a gene for expressing a reporter protein.
23 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses chitin synthase, (2) a gene that expresses chitosanase, and (3) a gene that expresses chitin deacetylase.
24 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses chitin synthase comprising SEQ ID NO. 2 or at least 70% homology thereto, (2) a gene that expresses chitosanase comprising SEQ ID NO. 3 or at least 70% homology thereto, and (3) a gene that expresses chitin deacetylase comprising SEQ ID NO. 4 or at least 70% homology thereto.
25 . The method of to claim 23 , wherein the construct comprises SEQ ID NO. 7 or at least 70% homology thereto, SEQ ID NO. 8 or at least 70% homology thereto, SEQ ID NO. 9, or at least 70% homology thereto, or SEQ ID NO. 10 or at least 70% homology thereto.
26 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses lipase, (2) a gene that expresses chitin synthase, (3) a gene that expresses chitosanase, and (4) a gene that expresses chitin deacetylase.
27 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses lipase comprising SEQ ID NO. 1 or at least 70% homology thereto, (2) a gene that expresses chitin synthase comprising SEQ ID NO. 2 or at least 70% homology thereto, (3) a gene that expresses chitosanase comprising SEQ ID NO. 3 or at least 70% homology thereto, and (4) a gene that expresses chitin deacetylase comprising SEQ ID NO. 4 or at least 70% homology thereto.
28 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses lipase, (2) a gene that expresses chitosanase, (3) a gene that expresses chitin synthase, and (4) a gene that expresses chitin deacetylase.
29 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses lipase comprising SEQ ID NO. 1 or at least 70% homology thereto, (2) a gene that expresses chitosanase comprising SEQ ID NO. 3 or at least 70% homology thereto, (3) a gene that expresses chitin synthase comprising SEQ ID NO. 2 or at least 70% homology thereto, and (4) a gene that expresses chitin deacetylase comprising SEQ ID NO. 4 or at least 70% homology thereto.
30 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a ribosomal binding site, (2) a gene that expresses lipase, (3) a ribosomal binding site, (4) a gene that expresses chitosanase, (5) a T7 promoter, (6) a ribosomal binding site, (7) a gene that expresses chitin synthase, (8) a ribosomal binding site, and (9) a gene that expresses chitin deacetylase.
31 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses lipase, (2) a gene that expresses chitin synthase, (3) a gene that expresses chitosanase, (4) a gene that expresses chitin deacetylase, and (5) a gene that expresses (1→3), (1→4)-β-glucan synthase.
32 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses lipase comprising SEQ ID NO. 1 or at least 70% homology thereto, (2) a gene that expresses chitin synthase comprising SEQ ID NO. 2 or at least 70% homology thereto, (3) a gene that expresses chitosanase comprising SEQ ID NO. 3 or at least 70% homology thereto, (4) a gene that expresses chitin deacetylase comprising SEQ ID NO. 4 or at least 70% homology thereto, and (5) a gene that expresses (1→3), (1→4)-β-glucan synthase comprising SEQ ID NO. 6 or at least 70% homology thereto.
33 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses chitin synthase, (2) a gene that expresses chitosanase, (3) a gene that expresses chitin deacetylase, and (4) a gene that expresses (1→3), (1→4)-β-glucan synthase.
34 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses chitin synthase comprising SEQ ID NO. 2 or at least 70% homology thereto, (2) a gene that expresses chitosanase comprising SEQ ID NO. 3 or at least 70% homology thereto, (3) a gene that expresses chitin deacetylase comprising SEQ ID NO. 4 or at least 70% homology thereto, and (4) a gene that expresses (1→3), (1→4)-β-glucan synthase comprising SEQ ID NO. 6 or at least 70% homology thereto.
35 . The method according to claim 1 , wherein the construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses chitin synthase, (2) a gene that expresses chitosanase, (3) a gene that expresses chitin deacetylase, (4) a gene that expresses (1→3), (1→4)-β-glucan synthase, (5) a gene that expresses transglycosylase, and (6) a gene that expresses chitin synthase regulatory factor CHR1.
36 . The method of claim 1 , further comprising contacting the plant with a second extract produced from a second biological device, wherein the second biological device comprises second host cells transformed with a supplemental DNA construct comprising the following genetic components: (a) a gene that expresses transglycosylase and (b) a gene that expresses chitin synthase regulatory factor CRH1.
37 . The method of claim 36 , wherein the supplemental DNA construct comprises from 5′ to 3′ the following genetic components in the following order: (1) a gene that expresses transglycosylase and (2) a gene that expresses chitin synthase regulatory factor CRH1.