Controlled hydrolysis of lactonic sophorolipids from Starmerella bombicola yields a previously undescribed sophorose analog that potently induces cellulase in Trichoderma reesei Rut-C30. Acid treatment of natural sophorolipids results in a mixture of monoacetylated, deacetylated, and diacetylated sophorolipids in acidic and lactonic forms. Isolation of the active components of the mixture, followed by structure determination by MS and NMR, reveals a new chemical entity, in which the lactone ring has been opened at the C-1' rather than at the C-4″ position of the sophorose moiety. This sophorose ester is resistant to degradation by the host and is at least 28 times more powerful an inducer than sophorose in shake-flask culture. Even at low concentrations (0.05 mM), the chemically modified sophorolipid effectively induces cellulase. With further improvements, this highly enabling technology can potentially reduce the cost of enzymes produced in T. reesei and can facilitate the rapid deployment of enzyme plants to support the nascent cellulosic biofuels and biochemicals industries.

gene expression, fermentation, cellulase, Trichoderma, sophorolipid, inducer

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

[*]O[C@@H]1[C@@H](COC(C)=O)O[C@@H](O[C@@H]2[C@@H](O)[C@H](O)[C@@H](COC(C)=O)O[C@H]2OC(C)CCCCCC/C=C\CCCCCCCC([*])=O)[C@H](O)[C@H]1O

688.808 g/mol (C₃₄H₅₆R₂O₁₄, R = next and previous repeats, not counted in mol weight)

Controlled hydrolysis of lactonic sophorolipids from Starmerella bombicola yields a previously undescribed sophorose analog that potently induces cellulase in Trichoderma reesei Rut-C30. Acid treatment of natural sophorolipids results in a mixture of monoacetylated, deacetylated, and diacetylated sophorolipids in acidic and lactonic forms. Isolation of the active components of the mixture, followed by structure determination by MS and NMR, reveals a new chemical entity, in which the lactone ring has been opened at the C-1' rather than at the C-4″ position of the sophorose moiety. This sophorose ester is resistant to degradation by the host and is at least 28 times more powerful an inducer than sophorose in shake-flask culture. Even at low concentrations (0.05 mM), the chemically modified sophorolipid effectively induces cellulase. With further improvements, this highly enabling technology can potentially reduce the cost of enzymes produced in T. reesei and can facilitate the rapid deployment of enzyme plants to support the nascent cellulosic biofuels and biochemicals industries.

gene expression, fermentation, cellulase, Trichoderma, sophorolipid, inducer

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

CC(=O)OC[C@H]1O[C@@H](O[C@@H]2[C@@H](O)[C@H](O)[C@@H](COC(C)=O)O[C@H]2OC(C)CCCCCC/C=C\CCCCCCCC(=O)O)[C@H](O)[C@@H](O)[C@@H]1O

706.823 g/mol (C₃₄H₅₈O₁₅)

Controlled hydrolysis of lactonic sophorolipids from Starmerella bombicola yields a previously undescribed sophorose analog that potently induces cellulase in Trichoderma reesei Rut-C30. Acid treatment of natural sophorolipids results in a mixture of monoacetylated, deacetylated, and diacetylated sophorolipids in acidic and lactonic forms. Isolation of the active components of the mixture, followed by structure determination by MS and NMR, reveals a new chemical entity, in which the lactone ring has been opened at the C-1' rather than at the C-4″ position of the sophorose moiety. This sophorose ester is resistant to degradation by the host and is at least 28 times more powerful an inducer than sophorose in shake-flask culture. Even at low concentrations (0.05 mM), the chemically modified sophorolipid effectively induces cellulase. With further improvements, this highly enabling technology can potentially reduce the cost of enzymes produced in T. reesei and can facilitate the rapid deployment of enzyme plants to support the nascent cellulosic biofuels and biochemicals industries.

gene expression, fermentation, cellulase, Trichoderma, sophorolipid, inducer

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6	C7	C8	C9	C10	C11	C12	C13	C14	C15	C16	C17	C18
2,4	l?17HOOle	172.3	33.7	24.6	25.6-29.6	25.6-29.6	25.6-29.6	25.6-29.6	26.9	129.6	129.6	26.9	25.6-29	25.6-29	25.6-29	25.6-29	39.0	71.5	22.2
2	bDGlcp	104.7	74.1	72.3	70.6	67.3	62.6
6	Ac	171.2	19.5
	bDGlcp	92.4	82.1	74.1	70.2	71.9	61.4


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6	H7	H8	H9	H10	H11	H12	H13	H14	H15	H16	H17	H18
2,4	l?17HOOle	-	2.36	1.62	1.27-1.46	1.27-1.46	1.27-1.46	1.27-1.46	2.00-2.05	5.34	5.34	2.00-2.05	1.27-1.46	1.27-1.46	1.27-1.46	1.27-1.46	1.40	3.78	1.14
2	bDGlcp	4.53	3.40	3.85	4.82	3.68	4.08-4.14
6	Ac	-	2.04
	bDGlcp	5.31	3.42	3.56	3.36	3.66	3.68-3.78


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6	C7/H7	C8/H8	C9/H9	C10/H10	C11/H11	C12/H12	C13/H13	C14/H14	C15/H15	C16/H16	C17/H17	C18/H18
2,4	l?17HOOle		33.7/2.36	24.6/1.62	25.6-29.6/1.27-1.46	25.6-29.6/1.27-1.46	25.6-29.6/1.27-1.46	25.6-29.6/1.27-1.46	26.9/2.00-2.05	129.6/5.34	129.6/5.34	26.9/2.00-2.05	25.6-29/1.27-1.46	25.6-29/1.27-1.46	25.6-29/1.27-1.46	25.6-29/1.27-1.46	39.0/1.40	71.5/3.78	22.2/1.14
2	bDGlcp	104.7/4.53	74.1/3.40	72.3/3.85	70.6/4.82	67.3/3.68	62.6/4.08-4.14
6	Ac		19.5/2.04
	bDGlcp	92.4/5.31	82.1/3.42	74.1/3.56	70.2/3.36	71.9/3.66	61.4/3.68-3.78

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

CC(=O)OC[C@H]1O[C@@H](O)[C@H](O[C@@H]2O[C@H](CO)[C@@H](OC(=O)CCCCCCC/C=C\CCCCCCC(C)O)[C@H](O)[C@H]2O)[C@@H](O)[C@@H]1O

664.786 g/mol (C₃₂H₅₆O₁₄)

Controlled hydrolysis of lactonic sophorolipids from Starmerella bombicola yields a previously undescribed sophorose analog that potently induces cellulase in Trichoderma reesei Rut-C30. Acid treatment of natural sophorolipids results in a mixture of monoacetylated, deacetylated, and diacetylated sophorolipids in acidic and lactonic forms. Isolation of the active components of the mixture, followed by structure determination by MS and NMR, reveals a new chemical entity, in which the lactone ring has been opened at the C-1' rather than at the C-4″ position of the sophorose moiety. This sophorose ester is resistant to degradation by the host and is at least 28 times more powerful an inducer than sophorose in shake-flask culture. Even at low concentrations (0.05 mM), the chemically modified sophorolipid effectively induces cellulase. With further improvements, this highly enabling technology can potentially reduce the cost of enzymes produced in T. reesei and can facilitate the rapid deployment of enzyme plants to support the nascent cellulosic biofuels and biochemicals industries.

gene expression, fermentation, cellulase, Trichoderma, sophorolipid, inducer

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

CC(O)CCCCCC/C=C\CCCCCCCC(=O)O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]2O)[C@H](O)[C@H]1O

622.749 g/mol (C₃₀H₅₄O₁₃)