Exo-endo cellulase fusion protein

Bower, Benjamin S; Larenas, Edmund A; Mitchinson, Colin

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Title: Exo-endo cellulase fusion protein

Abstract

The present invention relates to a heterologous exo-endo cellulase fusion construct, which encodes a fusion protein having cellulolytic activity comprising a catalytic domain derived from a fungal exo-cellobiohydrolase and a catalytic domain derived from an endoglucanase. The invention also relates to vectors and fungal host cells comprising the heterologous exo-endo cellulase fusion construct as well as methods for producing a cellulase fusion protein and enzymatic cellulase compositions.

Inventors:

Bower, Benjamin S ^[1]; Larenas, Edmund A ^[1]; Mitchinson, Colin ^[1]

Palo Alto, CA

Issue Date:: Tue Jan 17 00:00:00 EST 2012

Research Org.:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Org.:: USDOE

OSTI Identifier:: 1034913

Patent Number(s):: 8097445

Application Number:: 11/088,306

Assignee:: Danisco US Inc. (Palo Alto, CA)

Patent Classifications (CPCs):: C - CHEMISTRY C07 - ORGANIC CHEMISTRY C07K - PEPTIDES

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

Show more

C - CHEMISTRY C07 - ORGANIC CHEMISTRY C07K - PEPTIDES
C07K2319/00 - Fusion polypeptide
C07K2319/50 - containing protease site

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C12N15/625 - {containing a sequence coding for a signal sequence}
C12N15/80 - for fungi
C12N9/2437 - {Cellulases

C - CHEMISTRY C12 - BIOCHEMISTRY C12P - FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {
C12P21/02 - having a known sequence of two or more amino acids, e.g. glutathione
C12P21/06 - produced by the hydrolysis of a peptide bond, e.g. hydrolysate products

C - CHEMISTRY C12 - BIOCHEMISTRY C12Y - ENZYMES
C12Y302/01004 - Cellulase
C12Y302/01091 - Cellulose 1,4-beta-cellobiosidase

Show less

DOE Contract Number:: AC36-99GO10337

Resource Type:: Patent

Country of Publication:: United States

Language:: English

Subject:: 60 APPLIED LIFE SCIENCES

Citation Formats


                    Bower, Benjamin S, Larenas, Edmund A, and Mitchinson, Colin. Exo-endo cellulase fusion protein.  United States: N. p., 2012. 
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                    Bower, Benjamin S, Larenas, Edmund A, & Mitchinson, Colin. Exo-endo cellulase fusion protein.  United States.

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                    Bower, Benjamin S, Larenas, Edmund A, and Mitchinson, Colin. Tue .  
        "Exo-endo cellulase fusion protein".  United States.  https://www.osti.gov/servlets/purl/1034913.

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@article{osti_1034913,

  title        = {Exo-endo cellulase fusion protein},

  author       = {Bower, Benjamin S and Larenas, Edmund A and Mitchinson, Colin},

  abstractNote = {The present invention relates to a heterologous exo-endo cellulase fusion construct, which encodes a fusion protein having cellulolytic activity comprising a catalytic domain derived from a fungal exo-cellobiohydrolase and a catalytic domain derived from an endoglucanase. The invention also relates to vectors and fungal host cells comprising the heterologous exo-endo cellulase fusion construct as well as methods for producing a cellulase fusion protein and enzymatic cellulase compositions.},

  doi          = {},

  journal      = {},
number       = ,

  volume       = ,

  place        = {United States},

  year         = {Tue Jan 17 00:00:00 EST 2012},

  month        = {Tue Jan 17 00:00:00 EST 2012}

}

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Dawes, Iw
Biochemical Education, Vol. 11, Issue 2
https://doi.org/10.1016/0307-4412(83)90062-6

Isolation and Characterization of Acidothermus cellulolyticus gen. nov., sp. nov., a New Genus of Thermophilic, Acidophilic, Cellulolytic Bacteria
journal, July 1986

MOHAGHEGHI, A.; GROHMANN, K.; HIMMEL, M.
International Journal of Systematic Bacteriology, Vol. 36, Issue 3, p. 435-443
https://doi.org/10.1099/00207713-36-3-435

Cellulases of Trichoderma reesei
journal, January 1988

Schülein, Martin
Methods in Enzymology, Vol. 160, p. 234-242
https://doi.org/10.1016/0076-6879(88)60125-X

Engineering of a glycosidase Family 7 cellobiohydrolase to more alkaline pH optimum: the pH behaviour of Trichoderma reesei Cel7A and its E223S/ A224H/L225V/T226A/D262G mutant
journal, May 2001

Becker, Dieter; Braet, Christophe; Brumer Iii, Harry
Biochemical Journal, Vol. 356, Issue 1
https://doi.org/10.1042/0264-6021:3560019

Characterization of the secreted cellulases of Trichoderma reesei wild type and mutants during controlled fermentations
journal, July 1984

Sheir-Neiss, G.; Montenecourt, B. S.
Applied Microbiology and Biotechnology, Vol. 20, Issue 1, p. 46-53
https://doi.org/10.1007/BF00254645

Multiple roles of the cellulase CBHI in enhancing production of fusion antibodies by the filamentous fungus Trichoderma reesei
journal, June 1995

Nyyss�nen, Eini; Ker�nen, Sirkka
Current Genetics, Vol. 28, Issue 1
https://doi.org/10.1007/BF00311884

Structure of Cellulases and Their Applications
journal, April 1997

Ohmiya, Kunio; Sakka, Kazuo; Karita, Shuichi
Biotechnology and Genetic Engineering Reviews, Vol. 14, Issue 1, p. 365-414
https://doi.org/10.1080/02648725.1997.10647949

Amino acid substitution matrices from protein blocks.
journal, November 1992

Henikoff, S.; Henikoff, J. G.
Proceedings of the National Academy of Sciences, Vol. 89, Issue 22, p. 10915-10919
https://doi.org/10.1073/pnas.89.22.10915

Molecular evidence that the asexual industrial fungus Trichoderma reesei is a clonal derivative of the ascomycete Hypocrea jecorina.
journal, July 1996

Kuhls, K.; Lieckfeldt, E.; Samuels, G. J.
Proceedings of the National Academy of Sciences, Vol. 93, Issue 15, p. 7755-7760
https://doi.org/10.1073/pnas.93.15.7755

Expression of xylanase enzymes from thermophilic microorganisms in fungal hosts
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Bergquist, Peter; Te'o, Valentino; Gibbs, Moreland
Extremophiles, Vol. 6, Issue 3
https://doi.org/10.1007/s00792-001-0252-5

A versatile transformation system for the cellulolytic filamentous fungus Trichoderma reesei
journal, January 1987

Penttilä, Merja; Nevalainen, Helena; Rättö, Marjaana
Gene, Vol. 61, Issue 2, p. 155-164
https://doi.org/10.1016/0378-1119(87)90110-7

Enzymic activities of endo-1,4-β-d-glucanases purified from Trichoderma viride
journal, March 1978

Shoemaker, S. P.; Brown, R. D.
Biochimica et Biophysica Acta (BBA) - Enzymology, Vol. 523, Issue 1, p. 133-146
https://doi.org/10.1016/0005-2744(78)90016-5

Studies on the chromatographic fractionation of Trichoderma reesei cellulases by hydrophobic interaction
journal, December 1999

Tomaz, Cândida T.; Queiroz, João A.
Journal of Chromatography A, Vol. 865, Issue 1-2, p. 123-128
https://doi.org/10.1016/S0021-9673(99)00851-1

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Title: Exo-endo cellulase fusion protein

Abstract

Citation Formats

Ion-exchange chromatographic purification and quantitative analysis of Trichoderma reesei cellulases cellobiohydrolase I, II and endoglucanase II by fast protein liquid chromatography journal, May 1998

Studies of the cellulolytic system of Trichoderma reesei QM 9414. Analysis of domain function in two cellobiohydrolases by limited proteolysis journal, January 1988

A novel, small endoglucanase gene, egl5, from Trichoderma reesei isolated by expression in yeast journal, July 1994

Characteristics of fungal cellulases journal, January 1991

A bifunctional exoglucanase-endoglucanase fusion protein journal, January 1987

Molecular cloning and characterization of the glucoamylase gene of Aspergillus awamori. journal, November 1984

Separation of endo- and exo-type cellulases using a new affinity chromatography method journal, April 1984

Conversion of a β-Ketoacyl Synthase to a Malonyl Decarboxylase by Replacement of the Active-Site Cysteine with Glutamine journal, September 1999

Characterization of cellulases by HPLC separation journal, January 1983

Hydrolysis of cellulose using ternary mixtures of purified celluloses journal, March 1998

Characterization of a Thermomonospora fusca exocellulase journal, March 1995

CelS: a novel endoglucanase identified from Erwinia carotovora subsp. carotovora journal, May 1990

Dilute-Sulfuric Acid Pretreatment of Corn Stover in Pilot-Scale Reactor: Investigation of Yields, Kinetics, and Enzymatic Digestibilities of Solids journal, January 2003

Complete nucleotide sequence of a gene coding for Aspergillus aculeatus cellulase (FI-CMCase) journal, January 1990

Cloning and sequencing of cellulase cDNA from Aspergillus kawachii and its expression in Saccharomyces cerevisiae journal, April 1995

Purification and characterization of a β-glucosidase from solid-state cultures of Humicola grisea var. thermoidea journal, January 1996

Transformation of Trichoderma harzianum by high-voltage electric pulse journal, February 1990

Cloning and Expression of a Thermomonospora YX Endocellulase Gene in E. coli journal, September 1983

Homologous domains in Trichoderma reesei cellulolytic enzymes: Gene sequence and expression of cellobiohydrolase II journal, January 1987

ACEII, a Novel Transcriptional Activator Involved in Regulation of Cellulase and Xylanase Genes of Trichoderma reesei journal, April 2001

Antibodies specific for the human retinoblastoma protein identify a family of related polypeptides. journal, November 1991

Use of Aspergillus overproducing mutants, cured for intergrated plasmid, to overproduce heterologous proteins journal, August 1993

Purification and properties of an extracellular β-glucosidase from Lenzites trabea journal, January 1978

Fractionation of cellulase and β-glucosidase in a Trichoderma reesei culture liquid by use of two-phase partitioning journal, November 1999

The X-ray crystal structure of the Trichoderma reesei family 12 endoglucanase 3, Cel12A, at 1.9 Å resolution1 1Edited by A. R. Fersht journal, April 2001

Gapped BLAST and PSI-BLAST: a new generation of protein database search programs journal, September 1997

Properties of cellulolytic enzyme systems journal, April 1985

Improved tools for biological sequence comparison. journal, April 1988

Protein production by the filamentous fungus Trichoderma reesei : secretion of active antibody molecules journal, December 1995

Heterologous Protein Production by Filamentous Fungi journal, December 1991

Molecular Characterization and Heterologous Expression of the Gene Encoding a Low-Molecular-Mass Endoglucanase from Trichoderma reesei QM9414 journal, January 1998

Homology between cellulase genes of Trichoderma reesei: complete nucleotide sequence of the endoglucanase I gene journal, January 1986

Site-directed mutagenesis of the putative catalytic residues of Trichoderma reesci cellobiohydrolase I and endoglucanase I journal, November 1990

Specific quantification of trichoderma reesei cellulases in reconstituted mixtures and its application to cellulase-cellulose binding studies journal, October 1994

Limited proteolysis of the cellobiohydrolase I from Trichoderma reesei: Separation of functional domains journal, August 1986

Biolistic transformation of Trichoderma harzianum and Gliocladium virens using plasmid and genomic DNA journal, October 1993

Molecular Cloning of Exo–Cellobiohydrolase I Derived from Trichoderma Reesei Strain L27 journal, October 1983

Cloning and sequencing of the cDNA encoding β-glucosidase 1 from Aspergillus aculeatus journal, September 1996

Transformation of Aspergillus nidulans by using a trpC plasmid. journal, March 1984

Improved transformation efficiency of Aspergillus niger using the homologous niaD gene for nitrate reductase journal, July 1989

Two different types of intervening sequences in the glucoamylase gene from Aspergillus niger. journal, July 1984

[79] Purification of all glycolytic enzymes from one muscle extract journal, January 1982

Melamine Deaminase and Atrazine Chlorohydrolase: 98 Percent Identical but Functionally Different journal, April 2001

[57] Rethinking your purification procedure book, January 1990

Catalytic Plasticity of Fatty Acid Modification Enzymes Underlying Chemical Diversity of Plant Lipids journal, November 1998

cDNA Cloning of a Trichoderma reesei Cellulase and Demonstration of Endoglucanase Activity by Expression in Yeast journal, October 1997

Simultaneous saccharification and fermentation of lignocellulosic wastes to ethanol using a thermotolerant yeast journal, April 2001

Basic local alignment search tool journal, October 1990

EGIII, a new endoglucanase from Trichoderma reesei: the characterization of both gene and enzyme journal, March 1988

A general method applicable to the search for similarities in the amino acid sequence of two proteins journal, March 1970

Comparison of biosequences journal, December 1981

Trichoderma reesei cellobiohydrolase I with an endoglucanase cellulose-binding domain: action on bacterial microcrystalline cellulose journal, September 1997

Production of recombinant proteins in the filamentous fungus Trichoderma reesei journal, January 1995

Methods for measuring cellulase activities journal, January 1988

Cloning and relational analysis of 15 novel fungal endoglucanases from family 12 glycosyl hydrolase journal, May 2002

Efficient secretion of two fungal cellobiohydrolases by Saccharomyces cerevisiae journal, March 1988

Measurement of protein using bicinchoninic acid journal, October 1985

Cellulase families and their genes journal, September 1987

Genetic transformation of intact cells of Bacillus subtilis by electroporation journal, January 1990

Expression and processing of a major xylanase (XYN2) from the thermophilic fungus Humicola grisea var. thermoidea in Trichoderma reesei journal, February 2002

Analytical separation of Trichoderma reesei cellulases by ion-exchange fast protein liquid chromatography journal, January 1987

Rapid transformation of high cellulase-producing mutant strains of Trichoderma reesei by microprojectile bombardment journal, April 2000

The Molecular Biology of the Yeast Saccharomyces. Life Cycle and Inheritance journal, April 1983

Isolation and Characterization of Acidothermus cellulolyticus gen. nov., sp. nov., a New Genus of Thermophilic, Acidophilic, Cellulolytic Bacteria journal, July 1986

Cellulases of Trichoderma reesei journal, January 1988

Engineering of a glycosidase Family 7 cellobiohydrolase to more alkaline pH optimum: the pH behaviour of Trichoderma reesei Cel7A and its E223S/ A224H/L225V/T226A/D262G mutant journal, May 2001

Characterization of the secreted cellulases of Trichoderma reesei wild type and mutants during controlled fermentations journal, July 1984

Multiple roles of the cellulase CBHI in enhancing production of fusion antibodies by the filamentous fungus Trichoderma reesei journal, June 1995

Structure of Cellulases and Their Applications journal, April 1997

Amino acid substitution matrices from protein blocks. journal, November 1992

Molecular evidence that the asexual industrial fungus Trichoderma reesei is a clonal derivative of the ascomycete Hypocrea jecorina. journal, July 1996

Expression of xylanase enzymes from thermophilic microorganisms in fungal hosts journal, June 2002

A versatile transformation system for the cellulolytic filamentous fungus Trichoderma reesei journal, January 1987

Enzymic activities of endo-1,4-β-d-glucanases purified from Trichoderma viride journal, March 1978

Studies on the chromatographic fractionation of Trichoderma reesei cellulases by hydrophobic interaction journal, December 1999

Ion-exchange chromatographic purification and quantitative analysis of Trichoderma reesei cellulases cellobiohydrolase I, II and endoglucanase II by fast protein liquid chromatography
journal, May 1998

Studies of the cellulolytic system of Trichoderma reesei QM 9414. Analysis of domain function in two cellobiohydrolases by limited proteolysis
journal, January 1988

A novel, small endoglucanase gene, egl5, from Trichoderma reesei isolated by expression in yeast
journal, July 1994

Characteristics of fungal cellulases
journal, January 1991

A bifunctional exoglucanase-endoglucanase fusion protein
journal, January 1987

Molecular cloning and characterization of the glucoamylase gene of Aspergillus awamori.
journal, November 1984

Separation of endo- and exo-type cellulases using a new affinity chromatography method
journal, April 1984

Conversion of a β-Ketoacyl Synthase to a Malonyl Decarboxylase by Replacement of the Active-Site Cysteine with Glutamine
journal, September 1999

Characterization of cellulases by HPLC separation
journal, January 1983

Hydrolysis of cellulose using ternary mixtures of purified celluloses
journal, March 1998

Characterization of a Thermomonospora fusca exocellulase
journal, March 1995

CelS: a novel endoglucanase identified from Erwinia carotovora subsp. carotovora
journal, May 1990

Dilute-Sulfuric Acid Pretreatment of Corn Stover in Pilot-Scale Reactor: Investigation of Yields, Kinetics, and Enzymatic Digestibilities of Solids
journal, January 2003

Complete nucleotide sequence of a gene coding for Aspergillus aculeatus cellulase (FI-CMCase)
journal, January 1990

Cloning and sequencing of cellulase cDNA from Aspergillus kawachii and its expression in Saccharomyces cerevisiae
journal, April 1995

Purification and characterization of a β-glucosidase from solid-state cultures of Humicola grisea var. thermoidea
journal, January 1996

Transformation of Trichoderma harzianum by high-voltage electric pulse
journal, February 1990

Cloning and Expression of a Thermomonospora YX Endocellulase Gene in E. coli
journal, September 1983

Homologous domains in Trichoderma reesei cellulolytic enzymes: Gene sequence and expression of cellobiohydrolase II
journal, January 1987

ACEII, a Novel Transcriptional Activator Involved in Regulation of Cellulase and Xylanase Genes of Trichoderma reesei
journal, April 2001

Antibodies specific for the human retinoblastoma protein identify a family of related polypeptides.
journal, November 1991

Use of Aspergillus overproducing mutants, cured for intergrated plasmid, to overproduce heterologous proteins
journal, August 1993

Purification and properties of an extracellular β-glucosidase from Lenzites trabea
journal, January 1978

Fractionation of cellulase and β-glucosidase in a Trichoderma reesei culture liquid by use of two-phase partitioning
journal, November 1999

The X-ray crystal structure of the Trichoderma reesei family 12 endoglucanase 3, Cel12A, at 1.9 Å resolution1 1Edited by A. R. Fersht
journal, April 2001

Gapped BLAST and PSI-BLAST: a new generation of protein database search programs
journal, September 1997

Properties of cellulolytic enzyme systems
journal, April 1985

Improved tools for biological sequence comparison.
journal, April 1988

Protein production by the filamentous fungus Trichoderma reesei : secretion of active antibody molecules
journal, December 1995

Heterologous Protein Production by Filamentous Fungi
journal, December 1991

Molecular Characterization and Heterologous Expression of the Gene Encoding a Low-Molecular-Mass Endoglucanase from Trichoderma reesei QM9414
journal, January 1998

Homology between cellulase genes of Trichoderma reesei: complete nucleotide sequence of the endoglucanase I gene
journal, January 1986

Site-directed mutagenesis of the putative catalytic residues of Trichoderma reesci cellobiohydrolase I and endoglucanase I
journal, November 1990

Specific quantification of trichoderma reesei cellulases in reconstituted mixtures and its application to cellulase-cellulose binding studies
journal, October 1994

Limited proteolysis of the cellobiohydrolase I from Trichoderma reesei: Separation of functional domains
journal, August 1986

Biolistic transformation of Trichoderma harzianum and Gliocladium virens using plasmid and genomic DNA
journal, October 1993

Molecular Cloning of Exo–Cellobiohydrolase I Derived from Trichoderma Reesei Strain L27
journal, October 1983

Cloning and sequencing of the cDNA encoding β-glucosidase 1 from Aspergillus aculeatus
journal, September 1996

Transformation of Aspergillus nidulans by using a trpC plasmid.
journal, March 1984

Improved transformation efficiency of Aspergillus niger using the homologous niaD gene for nitrate reductase
journal, July 1989

Two different types of intervening sequences in the glucoamylase gene from Aspergillus niger.
journal, July 1984

[79] Purification of all glycolytic enzymes from one muscle extract
journal, January 1982

Melamine Deaminase and Atrazine Chlorohydrolase: 98 Percent Identical but Functionally Different
journal, April 2001

[57] Rethinking your purification procedure
book, January 1990

Catalytic Plasticity of Fatty Acid Modification Enzymes Underlying Chemical Diversity of Plant Lipids
journal, November 1998

cDNA Cloning of a Trichoderma reesei Cellulase and Demonstration of Endoglucanase Activity by Expression in Yeast
journal, October 1997

Simultaneous saccharification and fermentation of lignocellulosic wastes to ethanol using a thermotolerant yeast
journal, April 2001

Basic local alignment search tool
journal, October 1990

EGIII, a new endoglucanase from Trichoderma reesei: the characterization of both gene and enzyme
journal, March 1988

A general method applicable to the search for similarities in the amino acid sequence of two proteins
journal, March 1970

Comparison of biosequences
journal, December 1981

Trichoderma reesei cellobiohydrolase I with an endoglucanase cellulose-binding domain: action on bacterial microcrystalline cellulose
journal, September 1997

Production of recombinant proteins in the filamentous fungus Trichoderma reesei
journal, January 1995

Methods for measuring cellulase activities
journal, January 1988

Cloning and relational analysis of 15 novel fungal endoglucanases from family 12 glycosyl hydrolase
journal, May 2002

Efficient secretion of two fungal cellobiohydrolases by Saccharomyces cerevisiae
journal, March 1988

Measurement of protein using bicinchoninic acid
journal, October 1985

Cellulase families and their genes
journal, September 1987

Genetic transformation of intact cells of Bacillus subtilis by electroporation
journal, January 1990

Expression and processing of a major xylanase (XYN2) from the thermophilic fungus Humicola grisea var. thermoidea in Trichoderma reesei
journal, February 2002

Analytical separation of Trichoderma reesei cellulases by ion-exchange fast protein liquid chromatography
journal, January 1987

Rapid transformation of high cellulase-producing mutant strains of Trichoderma reesei by microprojectile bombardment
journal, April 2000

The Molecular Biology of the Yeast Saccharomyces. Life Cycle and Inheritance
journal, April 1983

Isolation and Characterization of Acidothermus cellulolyticus gen. nov., sp. nov., a New Genus of Thermophilic, Acidophilic, Cellulolytic Bacteria
journal, July 1986

Cellulases of Trichoderma reesei
journal, January 1988

Engineering of a glycosidase Family 7 cellobiohydrolase to more alkaline pH optimum: the pH behaviour of Trichoderma reesei Cel7A and its E223S/ A224H/L225V/T226A/D262G mutant
journal, May 2001

Characterization of the secreted cellulases of Trichoderma reesei wild type and mutants during controlled fermentations
journal, July 1984

Multiple roles of the cellulase CBHI in enhancing production of fusion antibodies by the filamentous fungus Trichoderma reesei
journal, June 1995

Structure of Cellulases and Their Applications
journal, April 1997

Amino acid substitution matrices from protein blocks.
journal, November 1992

Molecular evidence that the asexual industrial fungus Trichoderma reesei is a clonal derivative of the ascomycete Hypocrea jecorina.
journal, July 1996

Expression of xylanase enzymes from thermophilic microorganisms in fungal hosts
journal, June 2002

A versatile transformation system for the cellulolytic filamentous fungus Trichoderma reesei
journal, January 1987

Enzymic activities of endo-1,4-β-d-glucanases purified from Trichoderma viride
journal, March 1978

Studies on the chromatographic fractionation of Trichoderma reesei cellulases by hydrophobic interaction
journal, December 1999