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United States Patent [w]

Murphy et al.

[54] a-GALACTOSIDASE

[75] Inventors: Dennis Murphy, Paoli; John Reid, Bryn Mawr, both of Pa.

[73] Assignee: Diversa Corporation, San Diego, Calif.

[21] Appl. No.: 08/613,220 [22] Filed: Mar. 8, 1996

[51] Int. CI.6 C12N 15/09; C12N 15/56;

C12N 9/40

[52] U.S. CI 435/208; 435/320.1; 435/252.3;

435/325; 435/419; 536/23.2; 536/24.32

[58] Field of Search 435/208, 320.1,

435/252.3, 325, 419; 536/23.2, 24.32

[56] References Cited

PUBLICATIONS

Keller, et al., "Thermococcus alcaliphilus sp. nov., a new hyperthermophilic archaeum growing on polysulphide at

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US005958751A [ii] Patent Number: 5,958,751 [45] Date of Patent: Sep. 28,1999

alkaline pH.," Archives of Microbiology, 1995, vol. 164, pp. 390-395.

Sakka et al. Biosci. Biotech. Biochem. 57(2):273-277, 1993.

Genbank Database Record NID g3236129 "Pyrococcus horikoshii OT3 genomic DNA, 287001-544000 nt. Position" Jul. 25, 1998.

Primary Examiner—Robert A. Wax
Assistant Examiner—Bradley S. Mayhew
Attorney, Agent, or Firm—Fish & Richardson PC.

[57] ABSTRACT

A thermostable alpha-glycosidase derived from various Thermococcus, alcaliphilus AEDII12RA is disclosed. The enzymes are produced from native or recombinant host cells and can be utilized in the food processing industry.

15 Claims, 3 Drawing Sheets

FIG. 1

TTG AGA GCG CTC GTC TTT CAC GGC AAC CTC CAG TAT GCC GAA ATC CCA 48 Leu Arg Ala Leu Val Phe His Gly Asn Leu Gin Tyr Ala Glu lie Pro

5 10 15

AAG AGC GAA ATC CCA AAG GTC ATA GAG AAG GCA TAC ATC CCA GTC ATC 96
Lys Ser Glu lie Pro Lys Val lie Glu Lys Ala Tyr lie Pro Val lie

20 25 30

GAG ACA CTG ATT AAA GAA GAA ATT CCT TTT GGG CTC AAC ATA ACG GGC 14 4
Glu Thr Leu lie Lys Glu Glu lie Pro Phe Gly Leu Asn lie Thr Gly
35 40 45

TAT ACC TTA AAG TTC CTC CCG AAG GAT ATT ATA GAC CTC GTT AAA GGG 192
Tyr Thr Leu Lys Phe Leu Pro Lys Asp lie lie Asp Leu Val Lys Gly
50 55 60

GGC ATC GCG AGT GAC CTG ATA GAG ATA ATC GGA ACG AGC TAC ACG CAC 240
Gly lie Ala Ser Asp Leu lie Glu lie lie Gly Thr Ser Tyr Thr His
65 70 75 80

GCA ATA CTC CCC CTC CTC CCG CTT AGC AGA GTA GAA GCA CAA GTT CAG 288
Ala lie Leu Pro Leu Leu Pro Leu Ser Arg Val Glu Ala Gin Val Gin

85 90 95

AGA GAT AGG GAA GTT AAG GAA GAG CTC TTC GAG CTT TCT CCA AAG GGA 336
Arg Asp Arg Glu Val Lys Glu Glu Leu Phe Glu Val Ser Pro Lys Gly

100 105 110

TTC TGG CTG CCA GAG CTC GCC TAT GAC CCG ATA ATC CCT GCC ATA CTG 384
Phe Trp Leu Pro Glu Leu Ala Tyr Asp Pro lie lie Pro Ala lie Leu
115 120 125

[merged small][table]

FIG.1B

ATT GCA GGC TAC AGA ATG AGT GTT GAG GGA TTA TTA GAG GTT ATA GAC 816
lie Ala Gly Tyr Arg Met Ser Val Glu Gly Leu Leu Glu Val lie Asp

260 265 270

GAG CTC AAC TCG GAA CTG TGC CTT CCC TCA GAG CTG AAG CAC ATG GGA 8 64
Glu Leu Asn Ser Glu Leu Cys Leu Pro Ser Glu Leu Lys His Ser Gly
275 280 285

AGG GAG CTC TAC TTA CGG ACT TCG AGT TGG GCA CCA GAT AAG AGC TTG 912
Arg Glu Leu Tyr Leu Arg Thr Ser Ser Trp Ala Pro Asp Lys Ser Leu
290 295 300

AGG ATA TGG AGA GAG GAC GAA GGG AAC GCA AGA CTT AAT ATG CTG TCC 960
Arg lie Trp Arg Glu Asp Glu Gly Asn Ala Arg Leu Asn Met Leu Ser
305 310 315 320

TAC AAT ATG AGG GGC GAA CTC GCC CTT TTA GCC GAG AAC AGC GAT GCA 1008
Tyr Asn Met Arg Gly Glu Leu Ala Phe Leu Ala Glu Asn Ser Asp Ala

325 330 335

AGG GGA TGG GAG CCC CTC CCT GAG AGG AGG CTG GAT GCC TTC CGG GCG 1047
Arg Gly Trp Glu Pro Leu Pro Glu Arg Arg Leu Asp Ala Phe Arg Ala

340 345 350

ATA TAT AAC GAT TGG AGG GGT GAA AAT GGG GAA CCT TAG 108 6

lie Tyr Asn Asp Trp Arg Gly Glu Asn Gly Glu Pro End 355 360 365

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