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US4716115A Expired / Lapsed US ⏱️ 0.0 years remaining

Derived nisin producing microorganisms, method of production and use and products obtained thereby

Assignee: Indopco Inc
•
Filing: 1986-07-11
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Granted: 1987-12-29
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Expires: 2006-07-11
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Abstract

Abstract Derived microorganisms containing foreign DNA encoding for nisin production and a method for producing the derived microorganisms by transferring DNA to a recipient microorganism are described. The recipient microorganisms are preferably bacteria lacking in the ability to produce nisin. Nisin inhibits the growth of spoilage bacteria and is used in various materials for preservation, including foods and particularly at refrigeration temperatures. Nisin is also used in animals for improving the health of the animal. The foreign DNA is obtained from a donor microorganism and encodes for nisin production when transferred to the recipient microorganism.

Inventors

  • 👤 Carlos F. Gonzalez
  • 👤 Alfred J. Gryczka

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-09 10:18
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/195; C07K14/315; A; A23; A23K; A23K20/00; A23K20/10; A23K20/195; A23L; A23L13/00; A23L13/40; A23L13/45; C12; C12N; C12N1/00; C12N1/20; C12N1/205; C12P; C12P21/00; C12P21/02; A61; A61K; A61K38/00; C12R; C12R2001/00; C12R2001/01; C12R2001/46; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/885

Claims Analysis

Total Claims: 21
Claim 1
Independent Claim 95 words
A nisin producing engineered bacterium of the genus Streptococcus which contains foreign DNA which encodes for nisin production, which foreign DNA was obtained from a nisin producing donor bacterium and which foreign DNA was transferred to a recipient bacterium resulting in the nisin producing engineered bacterium, wherein the donor bacterium is a naturally occurring Streptococcus species containing plasmid DNA from a 29 Mdal plasmid encoding for sucrose utilization and nisin production and nisin resistance and wherein the recipient bacterium is a Streptococcus species which is sensitive to nisin and which does not naturally produce nisin.
Claim 2
Dependent Claim 95 words
A nisin producing engineered bacterium of the genus Streptococcus which contains foreign DNA which encodes for nisin production, which foreign DNA was obtained from a nisin producing donor bacterium and which foreign DNA was transferred to a recipient bacterium resulting in the nisin producing engineered bacterium, wherein the donor bacterium is a naturally occurring Streptococcus species containing plasmid DNA from a 29 Mdal plasmid encoding for sucrose utilization and nisin production and nisin resistance and wherein the recipient bacterium is a Streptococcus species which is sensitive to nisin and which does not naturally produce nisin.
Claim 3
Dependent Claim 27 words
The nisin producing engineered bacterium of claim 1 wherein the donor bacterium from which the plasmid DNA has been transferred is a nisin producing Streptococcus lactis strain.
Claim 4
Dependent Claim 32 words
The nisin producing engineered bacterium of claim 1 wherein the donor bacterium from which the plasmid DNA has been transferred is Streptococcus lactis and the recipient microorganism is Streptococcus lactis subspecies diacetilactis.
Claim 5
Dependent Claim 37 words
The nisin producing engineered bacterium of claim 1 wherein the donor from which the plasmid DNA has been transferred is selected from the group consisting of Streptococcus lactis ATCC 11,454, NRRL-B-15,458, NRRL-B-15,459, NRRL-B-15,460, NRRL-B-15,461, NRRL-B-15462 and NRRL-B-15470.
Claim 6
Dependent Claim 28 words
The nisin producing engineered bacterium of claim 1 wherein the recipient is selected from the group consisting of Streptococcus lactis subspecies diacetilactis NRRL-B-15005, NRRL-B-15006, NRRL-B-15018, NRRL-B-12070 and NRRL-B-12071.
Claim 7
Dependent Claim 65 words
The nisin producing engineered bacterium of claim 1 wherein the recipient bacterium to which the plasmid DNA has been transferred produces compounds inhibitory against Gram-negative bacteria, and in particular psychotrophic bacteria, and the donor bacterium from which the plasmid DNA has been transferred produces nisin so that nisin and the compounds inhibitory against the Gram-negative bacteria are both expressed by the engineered nisin producing bacterium.
Claim 8
Dependent Claim 33 words
The nisin producing engineered bacterium of claim 6 which is inhibitory against Gram-positive bacteria and in particular spore-forming Gram-positive bacteria and also is inhibitory against Gram-negative bacteria and in particular psychotrophic Gram-negative bacteria.
Claim 9
Dependent Claim 25 words
The nisin producing engineered bacterium of claim 1 selected from the group consisting of Streptococcus lactis subspecies diacetilactis NRRL-B-15464, NRRL-B-15,465, NRRL-B-15,466, NRRL-B-15,467, NRRL-B-15,468 and NRRL-B-15,469.
Claim 10
Dependent Claim 36 words
The nisin producing engineered bacterium of claim 1 mixed with non-nisin producing Streptococcus lactis subspecies diacetilactis which has been selected for resistance to nisin and production of an inhibitory substance which is inhibitory to Gram-negative bacteria.
Claim 11
Dependent Claim 10 words
The engineered bacterium of claim 1 which is phage resistant.
Claim 12
Independent Claim 114 words
The method for engineering a nisin producing bacterium of the genus Streptococcus which comprises: (a) transferring plasmid DNA which encodes for nisin production in a donor bacterium to a recipient bacterium to produce a nisin producing engineered bacterium, wherein the plasmid DNA is foreign to the recipient bacterium and encodes for nisin production and nisin resistance in the recipient bacterium wherein the donor bacteria is a Streptococcus species containing plasmid DNA derived from a 29 Mdal plasmid encoding for sucrose utilization, nisin production and nisin resistance and wherein the recipient bacterium is a Streptococcus species which is sensitive to nisin and does not naturally produce nisin; and (b) isolating the nisin producing engineered bacterium.
Claim 13
Independent Claim 114 words
The method for engineering a nisin producing bacterium of the genus Streptococcus which comprises: (a) transferring plasmid DNA which encodes for nisin production in a donor bacterium to a recipient bacterium to produce a nisin producing engineered bacterium, wherein the plasmid DNA is foreign to the recipient bacterium and encodes for nisin production and nisin resistance in the recipient bacterium wherein the donor bacteria is a Streptococcus species containing plasmid DNA derived from a 29 Mdal plasmid encoding for sucrose utilization, nisin production and nisin resistance and wherein the recipient bacterium is a Streptococcus species which is sensitive to nisin and does not naturally produce nisin; and (b) isolating the nisin producing engineered bacterium.
Claim 14
Dependent Claim 26 words
The method of claim 11 wherein the donor bacterium is selected from the group consisting of Streptococcus lactis ATCC 11,454, NRRL-B-15,458, NRRL-B-15,459, NRRL-B-15,460, NRRL-B-15,461, NRRL-B-15,462, NRRL-B-15,470.
Claim 15
Dependent Claim 25 words
The method of claim 11 wherein the recipient bacterium is selected from the group consisting of Streptococcus lactis subspecies diacetilactis NRRL-B-15005, NRRL-B-15006, NRRL-B-15018, NRRL-B-12070, NRRL-B-12071.
Claim 16
Dependent Claim 29 words
The method of claim 11 wherein the engineered bacterium is a transconjugant selected from the group consisting of Streptococcus lactis subspecies diacetilactis NRRL-B-15464, NRRL-B-15,465, NRRL-B-15,466, NRRL-B-15,467, NRRL-B-15,468 and NRRL-B-15,469.
Claim 17
Dependent Claim 72 words
The bacterial composition which comprises in admixture: (a) a nisin producing Streptococcus containing foreign plasmid DNA from a 29 Mdal plasmid encoding for sucrose utilization and nisin production in a Streptococcus which is sensitive to nisin and does not naturally produce nisin; and (b) a nisin resistant Streptococcus diacetilactis which produces an inhibitory substance for Gram-negative bacteria, wherein the mixture of the bacteria in the composition produces the inhibitory substance and nisin.
Claim 18
Dependent Claim 72 words
The bacterial composition which comprises in admixture: (a) a nisin producing Streptococcus containing foreign plasmid DNA from a 29 Mdal plasmid encoding for sucrose utilization and nisin production in a Streptococcus which is sensitive to nisin and does not naturally produce nisin; and (b) a nisin resistant Streptococcus diacetilactis which produces an inhibitory substance for Gram-negative bacteria, wherein the mixture of the bacteria in the composition produces the inhibitory substance and nisin.
Claim 19
Dependent Claim 32 words
The composition of claim 15 containing at least 10 6 cells per ml as a frozen, dried or lyophilized live culture as a preserved form for storage and shipment prior to use.
Claim 20
Dependent Claim 22 words
The composition of claim 15 containing at least 10 9 cells per ml in a preserved form for storage prior to use.
Claim 21
Dependent Claim 16 words
The composition of claim 15 wherein the Streptococcus lactis subsp. diacetilactis is naturally resistant to nisin.

References Cited (Backward Citations)

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USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1990-12-28 1991-06-29 1991-12-29 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1994-12-28 1995-06-29 1995-12-29 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1998-12-28 1999-06-29 1999-12-29 $7700.00 Window Closed / Lapsed

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