IP Library Granted Patent US 12,215,367
Granted Patent B2
US 12,215,367 · App. 17/986,618 · Granted Feb 4, 2025

Lactase enzymes with improved properties

Inventors: Hans Raj (Hoersholm, DK); Pernille Smith (Broenshoej, DK); Thomas Eckhardt (Birkeroed, DK); Vojislav Vojinovic (Graested, DK); Charlotte Elisabeth Grüner Schöller (Virum, DK); Johannes Maarten van den Brink (Herlev, DK)
Assignee: Kerry Group Services International Ltd
C12N9/2471A23L29/06
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,215,367
App. No.
17/986,618
Granted
Feb 4, 2025
Kind
B2
Abstract

Disclosed are peptides exhibiting betagalactosidase enzyme activity as well as dairy products comprising them and related methods for reducing the lactose content in compositions, such as dairy products.

Claims (20)

1. A dairy product comprising a milk-based substrate and a peptide exhibiting beta-galactosidase enzyme activity, wherein the amino acid sequence of the peptide is selected from SEQ ID NO: 9, 10, 11, and 16, enzymatically active fragments thereof, and amino acid sequences having not more than 22 amino acid substitutions, additions or deletions relative to the amino acid sequence selected from SEQ ID NO: 9, 10, 11, and 16, and enzymatically active fragments thereof.

2. A method for producing a dairy product according to claim 1 , comprising:

adding the peptide exhibiting beta-galactosidase activity to a milk-based substrate comprising lactose; and

treating said milk-based substrate with said peptide.

3. The method according to claim 2 , wherein said treating takes place at a pH within a range of 3-10.

4. The method according to claim 2 , wherein all or a part of said treating takes place at a temperature of not more than 25° C.

5. The method according to claim 2 , wherein all or a part of said treating takes place at a temperature of at least 25° C.

6. The method according to claim 2 , wherein the galactose inhibition of said peptide or dimeric peptide is less than 60%.

7. The method according to claim 2 , wherein said dairy product is a fermented milk product and said adding is performed during or prior to fermentation.

8. The method according to claim 7 , which method does not require addition of a further enzyme after fermentation.

9. The method according to claim 2 , wherein said dairy product is a fermented milk product and said adding is performed immediately following fermentation.

10. The method according to claim 2 , wherein said dairy product is fresh milk and said adding is performed prior to, in conjunction with, or immediately following a step of pasteurization.

11. The method according to claim 2 , wherein said dairy product is ultra-heat treatment (UHT) milk and said adding is performed prior to, in conjunction with, or immediately following a step of ultra-heat treatment.

12. The method according to claim 2 , wherein said treating is started at a temperature of between 40° C. and 100° C.

13. The dairy product according to claim 1 , wherein the amino acid sequence of the peptide is selected from SEQ ID NO: 9, 10, 11, and 16, and amino acid sequences having not more than 22 amino acid substitutions, additions, or deletions relative to the amino acid sequence selected from SEQ ID NO: 9, 10, 11, and 16.

14. The dairy product according to claim 1 , wherein the peptide has the amino acid sequence of SEQ ID NO:9.

15. The dairy product according to claim 1 , wherein the peptide has the amino acid sequence of SEQ ID NO:10.

16. The dairy product according to claim 1 , wherein the peptide has the amino acid sequence of SEQ ID NO:11.

17. The dairy product according to claim 1 , wherein the peptide has the amino acid sequence of SEQ ID NO: 16.

18. The dairy product according to claim 1 , wherein dairy product is selected from lactose-free milk, low-lactose milk, yogurt, cheese, fermented milk products, dietary supplements, and probiotic dietary products.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 3, 2024
From: CHR. HANSEN A/S
To: KERRY GROUP SERVICES INTERNATIONAL LTD
Reel/Frame 067309/0489 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 14, 2022
From: RAJ, HANS; SMITH, PERNILLE; ECKHARDT, THOMAS; VOJINOVIC, VOJISLAV; SCHÖLLER, CHARLOTTE ELISABETH GRÜNER; VAN DEN BRINK, JOHANNES MAARTEN
To: CHR. HANSEN A/S
Reel/Frame 061763/0038 →
Priority Claims (1)
EP 17166021 · Apr 11, 2017 · regional
Continuity (2)
Continuation 16604129
Related Publication 20230235307A1 · Jul 27, 2023
References Cited (138)
US 8030049B2 · Tzortzis · 2011 [cited by examiner]
US 10058107B2 · Hendriksen et al. · 2018 [cited by applicant]
US 10306902B2 · Hendriksen et al. · 2019 [cited by applicant]
US 10555541B2 · Hendriksen et al. · 2020 [cited by applicant]
US 20090110770A1 · Tzortzis et al. · 2009 [cited by applicant]
US 20090117080A1 · Tzortzis et al. · 2009 [cited by applicant]
US 20090297660A1 · Silver et al. · 2009 [cited by applicant]
US 20100113383A1 · Mills et al. · 2010 [cited by applicant]
US 20100285175A1 · Hendriksen et al. · 2010 [cited by applicant]
US 20120058223A1 · Stougaard et al. · 2012 [cited by applicant]
US 20160333331A1 · De Jong et al. · 2016 [cited by applicant]
US 20170215449A1 · Nagahata et al. · 2017 [cited by applicant]
US 20190343138A1 · Ba et al. · 2019 [cited by applicant]
US 20200120946A1 · Hendriksen et al. · 2020 [cited by applicant]
US 20200123519A1 · Bongiorni et al. · 2020 [cited by applicant]
US 20210032615A1 · Raj et al. · 2021 [cited by applicant]
US 20210037844A1 · Hendriksen et al. · 2021 [cited by applicant]
US 20210348147A1 · Raj et al. · 2021 [cited by applicant]
US 20230076578A1 · Raj et al. · 2023 [cited by applicant]
US 20230210121A1 · Raj et al. · 2023 [cited by applicant]
CN 103431042B · 2015 [cited by applicant]
EP 1227152A1 · 2002 [cited by applicant]
EP 2530148A1 · 2012 [cited by applicant]
EP 2957180B1 · 2015 [cited by applicant]
RU 2278160C2 · 2005 [cited by applicant]
RU 2009120742 · 2010 [cited by applicant]
WO WO2005084411A2 · 2005 [cited by applicant]
WO WO2005086794A2 · 2005 [cited by applicant]
WO WO2007088324A1 · 2007 [cited by applicant]
WO WO2007110619A1 · 2007 [cited by applicant]
WO WO2008033520A2 · 2008 [cited by applicant]
WO WO2009009142A2 · 2009 [cited by applicant]
WO WO2009071539A1 · 2009 [cited by applicant]
WO WO2010092057A1 · 2010 [cited by applicant]
WO WO2013160413A1 · 2013 [cited by applicant]
WO WO2015107050A1 · 2015 [cited by applicant]
WO WO2017216000A1 · 2017 [cited by applicant]
WO WO2018041869A1 · 2018 [cited by applicant]
WO WO2018130630A1 · 2018 [cited by applicant]
WO WO2018187524A1 · 2018 [cited by applicant]
WO WO2018189238A1 · 2018 [cited by applicant]
WO WO2018189242A1 · 2018 [cited by applicant]
Odamaki et al., Comparative Genomics Revealed Genetic Diversity and Species/Strain-Level Differences in Carbohydrate Metabolism of Three Probiotic Bifidobacterial Species. Int. J. Genomics., 2015, Article ID 567809, 12 … [cited by examiner]
“Beta-galactosidase [ [cited by applicant]
“Beta-galactosidase [ [cited by applicant]
“Beta-galactosidase [ [cited by applicant]
“Beta-galactosidase [ [cited by applicant]
“Beta-galactosidase [ [cited by applicant]
U.S. Appl. No. 16/604,133, filed Oct. 9, 2019. [cited by applicant]
U.S. Appl. No. 16/604,134, filed Oct. 9, 2019. [cited by applicant]
“Chapter 3 Lactose content of milk and milk products,” The American Journal of Clinical Nutrition, vol. 48, No. 4 pp. 1099-1044 (Oct. 1988) Available online, URL: https://academic.oup.com/ajcn/article-abstract/48/4/1099… [cited by applicant]
“Uniprot: A0A0B5J47” (Apr. 1, 2015), Retrieved from the Internet, URL:http://ibis/exam/dbfetch.jsp?id=UNIPROT:A0A0B5J47 (Retrieved on May 11, 2017). [cited by applicant]
“Uniprot: A0AS2MCC8—beta galactosidase,” (Feb. 17, 2016) Retrieved from the Internet, URL: https://ibis/exam/dbfetch.jsp?id=UNIPROT:A0A0S2MCC8 [retrieved on Mar. 9, 2018). [cited by applicant]
Broune et al. “Catalytic Plasticity of Fatty Acid Modification Enzymes Underlying Chemical Diversity of Plant Lipids,” Science, vol. 282, pp. 1315-1317 (1998). [cited by applicant]
Database GenBank: ACE06986.1, (Jun. 8, 2012). [cited by applicant]
Database GenBank: CDR82630.1, (Jun. 11, 2014). [cited by applicant]
Devos et al., “Practical Limits of Function Prediction,” Proteins: Structure, Function, and Genetics, vol. 41, pp. 98-107 (Aug. 2000). [cited by applicant]
GenBank Accession No. CAI98003.1 Feb. 27, 2015. [cited by applicant]
Guo et al., “Protein tolerance to random amino acid change,” PNAS, vol. 101, No. 25, pp. 9205-9210 (Jun. 2004). [cited by applicant]
Horner et al., “ß-Galactosidase activity of commercial lactase samples in raw and pasteurized milk at refrigerated temperatures,” J. Dairy Sci. 94: 3242-3249 (2011). [cited by applicant]
Keskin et al., “A new, structurally nonredundant, diverse data set of protein-protein interfaces and its implications,” Protein Science, vol. 13, pp. 1043-1055 (2004). [cited by applicant]
Klimova E.V. Advantages of using beta-galactosidase for hydrolysis of lactose and obtaining galactooligosaccharides; prospects for the use of the obtained products in industrial food technologies, Food and processing in… [cited by applicant]
Kreft et al., “Lactose hydrolysing ability of sonicated cultures of [cited by applicant]
Nakagawa et al., “Overexpression and functional analysis of cold-active B-galactosidase from Arthrobacter psychrolocatohilus strain F2,” Protein Expression and Purification 54 (2007) 295-299 (Available on line Mar. 2007… [cited by applicant]
Office Action and Search Report issued on May 14, 2021 in Russian Application No. 2019134223/10. [cited by applicant]
Office Action issued on Apr. 12, 2022 in U.S. Appl. No. 16/998,706 (US 2021-0032615). [cited by applicant]
Office Action issued on Jan. 22, 2021, in U.S. Appl. No. 16/998,706 (US 2021-0032615). [cited by applicant]
Ogurtsov A.N., Methods of bioinformatic analysis, Textbook, Kharkov, 2011, NTU “KhPI”, pp. 4-5, 25. [cited by applicant]
Palak-Szukalska et al., “A novel cold-active B-D-galactosidase with transglycosylation activity from the [cited by applicant]
Rhimi et al., “Exploring the acidotolerance of ß-galactosidase from [cited by applicant]
Schmidt et al., “Identification, cloning and expression of a cold-active B-galactosidase from a novel Arctic bacterium, Alkalilactibacillus ikkense,” (2010) Environmental Technology, 31:10, 1107-1114 (Published online J… [cited by applicant]
Seffernick et al., “Melamine Deaminase and Atrazine Chlorohydrolase: 98 Percent Identical but Functionally Different,” Journal of Bacteriology, vol. 183, No. 8, pp. 2405-2410 (Apr. 2001). [cited by applicant]
Singer et al., “Genes & Genomes, A changing Perspective,” University Science Books Mill Valley, CA (1998). [cited by applicant]
U.S. Patent and Trademark Office; Final Office Action; U.S. Appl. No. 16/998,706; dated Jun. 9, 2021; 15 pages. [cited by applicant]
UniProt Accession No. A0AOS2MCC8 Feb. 17, 2016. [cited by applicant]
UniProt Accession No. F0K2P6, May 3, 2011. [cited by applicant]
UniProt Accession No. G6F860, Jan. 25, 2012. [cited by applicant]
UniProt Accession Nos. TrEMBL, A7A6G3_BIFAD, Sep. 11, 2007, Q38UW6_LACSS, Nov. 22, 2005, Q38UW7_LACSS, Nov. 22, 2005, R5YYAO_9LACO, Jul. 24, 2013, F0TG79_LACAM, May 3, 2011, K2MWD3_BIFBI, Nov. 28, 2012, D4QFE8_BIFBI, Ju… [cited by applicant]
Uniprot:G6F860 (Oct. 2020). [cited by applicant]
UniProtKB—A0A076JKA5 (A0A076JKA5_BIFAD); Oct. 29, 2014; 7 pages. [cited by applicant]
UniProtKB—A0A0A1GLP4 (A0A0A1GLP4_BIFLN); Feb. 4, 2015; 8 pages. [cited by applicant]
UniProtKB—A0A0A715K5 (A0A0A715K5_9BIFI); Mar. 4, 2015; 8 pages. [cited by applicant]
UniProtKB—A0A0H2P357 (A0A0H2P357_BIFBI); Sep. 16, 2015; 7 pages. [cited by applicant]
UniProtKB—A0A0U5FVZ6 (A0A0U5FVZ6_LACDE); Mar. 16, 2016; 9 pages. [cited by applicant]
UniProtKB—A0A126SWK6 (A0A126SWK6_9BIFI); Jul. 6, 2016; 8 pages. [cited by applicant]
UniProtKB—A0A174BAQ4 (A0A174BAQ4_9BIFI); Sep. 7, 2016; 8 pages. [cited by applicant]
UniProtKB—A0A174BB61 (A0A174BB61_BIFAD); Sep. 7, 2016; 8 pages. [cited by applicant]
UniProtKB—A0A174BH17 (A0A174BH17_9FIRM); Sep. 7, 2016; 5 pages. [cited by applicant]
UniProtKB—A0A1D7UM07 (A0A1D7UM07_BIFLN); Jan. 18, 2017; 8 pages. [cited by applicant]
UniProtKB—A0A1D7ZXL7 (A0A1D7ZXL7_LIMFE); Jan. 18, 2017; 7 pages. [cited by applicant]
UniProtKB—A0A1S2W2V3 (A0A1S2W2V3_BIFLN); Apr. 12, 2017; 8 pages. [cited by applicant]
UniProtKB—A0A1X2Z956 (A0A1X2Z956_BIFAD); Jul. 5, 2017; 8 pages. [cited by applicant]
UniProtKB—A0A1X2ZA47 (A0A1X2ZA47_BIFAD); Jul. 5, 2017; 7 pages. [cited by applicant]
UniProtKB—A0A1X2ZAP4 (A0A1X2ZAP4_BIFAD); Jul. 5, 2017; 7 pages. [cited by applicant]
UniProtKB—A0A2G5Q4A6 (A0A2G5Q4A6_9BIFI); Jan. 31, 2018; 8 pages. [cited by applicant]
UniProtKB—A0A4ROSL12 (A0A4ROSL12_BIFLN); Jul. 31, 2019; 8 pages. [cited by applicant]
UniProtKB—A0A4R0U1N4 (A0A4R0U1N4_BIFLN); Jul. 31, 2019; 8 pages. [cited by applicant]
UniProtKB—A0A6A2R535 (A0A6A2R535_BIFAD); Jun. 17, 2020; 7 pages. [cited by applicant]
UniProtKB—A0A6B1X5Q7 (A0A6B1X5Q7_9BIFI); Jun. 17, 2020; 6 pages. [cited by applicant]
UniProtKB—A0A611DQE1 (A0A611DQE1_BIFLN); Aug. 12, 2020; 8 pages. [cited by applicant]
UniProtKB—A0A6L4K944 (A0A6L4K944_BIFAD); Oct. 7, 2020; 7 pages. [cited by applicant]
UniProtKB—A0A6L4V5B5 (A0A6L4V5B5_9BIFI); Oct. 7, 2020; 8 pages. [cited by applicant]
UniProtKB—A0A7D9N5G4 (A0A7D9N5G4_LACJH); Dec. 2, 2020; 8 pages. [cited by applicant]
UniProtKB—A0A829LWJ6 (A0A829LWJ6_LIMFE); Sep. 29, 2021; 7 pages. [cited by applicant]
UniProtKB—A5VKG8 (A5VKG8_LIMRD); Jul. 10, 2007; 8 pages. [cited by applicant]
UniProtKB—B2GAA1 (B2GAA1_LIMF3); Jun. 10, 2008; 7 pages. [cited by applicant]
UniProtKB—B2GAA2 (B2GAA2_LIMF3); Jun. 10, 2008; 6 pages. [cited by applicant]
UniProtKB—D6ZY97 (D6ZY97_BIFLJ); Aug. 10, 2010; 8 pages. [cited by applicant]
UniProtKB—E4SLB1 (E4SLB1_LACAR); Feb. 8, 2011; 8 pages. [cited by applicant]
UniProtKB—E8MRV2 (E8MRV2_BIFL1); Apr. 5, 2011; 8 pages. [cited by applicant]
UniProtKB—F0HTF8 (F0HTF8_LACDL); May 3, 2011; 9 pages. [cited by applicant]
UniProtKB—F0TG75 (F0TG75_LACAM); May 3, 2011; 8 pages. [cited by applicant]
UniProtKB—F2M1D8 (F2MID8_LACAL); May 11, 2011; 8 pages. [cited by applicant]
UniProtKB—F4AFP0 (F4AFP0_LACJH); Jun. 28, 2011; 8 pages. [cited by applicant]
UniProtKB—F8ASA8 (F8ASA8_BIFLN); Sep. 21, 2011; 8 pages. [cited by applicant]
UniProtKB—G6F860 (G6F860_LACDE); Jan. 25, 2012; 9 pages. [cited by applicant]
UniProtKB—I3WJ66 (I3WJ66_BIFBI); Sep. 5, 2012; 8 pages. [cited by applicant]
UniProtKB—K215J0 (K215J0_BIFBI); Nov. 28, 2012; 8 pages. [cited by applicant]
UniProtKB—Q5FJD5 (Q5FJD5_LACAC); Mar. 1, 2005; 9 pages. [cited by applicant]
UniProtKB—Q74KL4 (Q74KL4_LACJO); Jul. 5, 2004; 8 pages. [cited by applicant]
UnitProtKB—D9ZDZ1 (D9ZDZ1_9ZZZZ); Oct. 5, 2010; 7 pages. [cited by applicant]
Van De Guchte, et al., Beta-galactosidase [ [cited by applicant]
Wang et al., “A novel cold-adapted B-galactosidase isolated from [cited by applicant]
Whisstock et al., “Prediction of protein function from protein sequence and structure,” Quarterly Reviews of Biophysics, vol. 36, No. 3 (pp. 307-340) (2003). [cited by applicant]
Wierzbicka-Wos et al., “A novel cold-active B-D-galactosidase from the [cited by applicant]
Witkowski et al. “Conversion of B-Ketoacyl Synthase to a Malonyl Decarboxylase by Replacement of the Active-Site Cysteine with Glutamine,” Biochemistry, vol. 38, pp. 11643-11650 (1999). [cited by applicant]
Skripnyuk A.A., et al.; “Modern methods for producing ß-galactosidase”; Science Innovations Technologies, 3; 2014; pp. 198-204. [cited by applicant]
Banerjee, Goutam et al.; “Is divalent magnesium cation the best cofactor for bacterial beta-galactosidase?”; J Biosci, vol. 43, No. 5; Oct. 4, 2018; pp. 941-945. [cited by applicant]
Biocceleration Ltd.; Seq Alignment Result (U.S. Appl. No. 16/604,134 SEQ #7 vs Tzortis et al. (US2009/0110770); Seq #2 using SLIC and ABSS SEQ—to SEQ (aa); Jun. 27, 2023. [cited by applicant]
Karlsson, Maria A. et al.; “Changes in stability and shelf-life of ultra-high temperature treated milk during long term storage at different temperatures”; Heliyon, vol. 5; Sep. 12, 2019; 9 pages. [cited by applicant]
Kuznetsova, E., “Brackets in Text of Legal Document as a Linguistic and Cognitive Phenomenon”; Institute of Humanities, Severodvinsk branch of Lomonosov Northern (Arctic) Federal University; Vestnik Moskovskogo gosudars… [cited by applicant]
Nguyen, Thao Thi et al.; “Effect of mutations to amino acid A301 and F361 in thermostability and catalytic activity of the beta-galactosidase from Bacillus subtilis VTCC-DVN-12-01”; BMC Biochemistry (2016) 17:15; Jul. 2… [cited by applicant]
Patent Office of the Russian Federation: Federal Institute of Industrial Property; Office Action (Enquiry) (English translation); Russian Patent Application No. 2021112325/10(026315); Jun. 19, 2023; 10 pages. [cited by applicant]
Seffernick, Jennifer et al.; “Melamine Deaminase and Atrazine Chlorohydrolase: 98 Percent Identical but Functionally Different”; Journal of Bacteriology, vol. 183, No. 8; Apr. 2001; pp. 2405-2410. [cited by applicant]
Whisstock, James C. et al.; “Prediction of protein function from protein sequence and structure”; Quarterly Review of Biophysics 36, 3; Aug. 2003; pp. 307-340. [cited by applicant]
Cecchini et al.; OM protein—protein search, using sw model; GenCore version 6.4.2; run on Jan. 31, 2022; title: US-16-604-129A-22; 6 pages. [cited by applicant]
Genbank Accession KRO12099 (https://www.ncbi.nlm.nih.gov/protein/KRO12099.1?report=genbank&log$=protalign&blast_rank=1&RID=NRM8Y7MN016); publication date: Nov. 6, 2015; 2 pages. [cited by applicant]
UniProt—A0A174B8K1_BIFAD; Sep. 7, 2016; 6 pages. [cited by applicant]
Cited By (1)
US 12,538,929