Antihypertensive peptides derived from caseins by proteolytic action.
1.Introduction
Peptides are well known as nitrogen sources to supply various amino acids for many different organisms, and also have many hormonal functions in our body. Previous studies have reported a secondary role for peptides with specific amino acid sequences that possess biological function
Among these bioactive peptides, antihypertensive peptides have been extensively studied and reviewed (5-8). Hypertension is a major risk factor in cardiovascular disease, such as heart disease and stroke. In order to reduce the incidence of disease, pharmacological substances can be used to decrease high blood pressure to within the normal range. Angiotensin I-converting enzyme (kininase II; EC 3.4.15.1) (ACE) is predominantly expressed as a membrane-bound ectoenzyme in vascular endothelial cells and several other cell types, including absorptive epithelia, neuroepithelia and male germinal cells (9, 10). A dipeptidyl carboxypeptidase, ACE catalyzes the production of a vasoconstrictor, angiotensin II, and inactivates a vasodilator, bradykinin (11, 12). The first competitive inhibitors to ACE were reported as naturally occurring peptides isolated from snake venom (13, 14). Then, inhibitory activities on ACE, which plays an important role in blood pressure regulation are generally assessed for preparation of antihypertensive peptides.
Among lactic acid bacteria
1.1. Antihypertensive peptide in fermented milk
1.1.1. ACE inhibitory peptides from milk proteins
Many kinds of ACE inhibitory (ACEI) peptides have been reported from enzymatic hydrolyzates of milk protein, as well as synthetic peptides and fermented products (16-24) (Table 1). Spontaneously hypertensive rat (SHR) is a useful animal model to evaluate the antihypertensive activity of ACEI peptides because the systolic blood pressure of SHR reaches over 230 mmHg and is powerful tool for detection of the
Peptide | Source | Preparation | (IC50(µM) | Dose (mg/kg) | (((SBP(mm Hg) | ||||||||||||
<Enzymatic hydrolysate"/ | |||||||||||||||||
FFVAPFPEVFGK | αs1-casein | Trypsin | 77 | 100 | -13.0 | ||||||||||||
AVPYPQR | -casein | Trypsin | 15 | 100 | -10.0 | ||||||||||||
TTMPLW | βs1-casein | Trypsin | 16 | 100 | -13.6 | ||||||||||||
LKPNM | Aldolase | Thermolysin | 2.4 | 60 | -23 | ||||||||||||
LKP | Aldolase | Chicken muscle | 0.32 | 60 | -18 | ||||||||||||
IPA | -lactogloblin | Proteinase K | 141 | 8 | -31 | ||||||||||||
VYPFPG | -casein | Proteinase K | 221 | 8 | -22 | ||||||||||||
GKP | -microglobulin | Proteinase K | 352 | 8 | -26 | ||||||||||||
FP | -casein, albumin | Proteinase K | 315 | 8 | -27 | ||||||||||||
YKVPQL | αs1-casein | Proteinase | 22 | 1 | -12.5 | ||||||||||||
<Fermented products"/ | |||||||||||||||||
RF | Sake lees | Brewing | ND | 100 | -17 | ||||||||||||
VW | Sake lees | Brewing | 1.4 | 100 | -10 | ||||||||||||
YW | Sake lees | Brewing | 10.5 | 100 | -28 | ||||||||||||
VY | Sake | Brewing | 7.1 | 100 | -31 | ||||||||||||
IYPRY | Sake | Brewing | 4.1 | 100 | -19 | ||||||||||||
VPP | -casein | Fermentation | 9 | 1.6 | -20 | ||||||||||||
IPP | - and κ-casein | Fermentation | 5 | 1 | -15.1 | ||||||||||||
YP | βs1, - and κ-casein | Fermentation | 720 | 1 | -27.4 | ||||||||||||
ND: Not described | |||||||||||||||||
(IC50: Peptide concentration that shows 50% inhibition of ACE activity | |||||||||||||||||
(((SBP: systolic blood pressure of spontaneously hypertensive rat |
Lactic acid bacteria have proteolytic systems that can hydrolyze milk protein and have been reported to utilize the peptides released from the milk protein casein (25-27). Among lactic acid bacteria
Strain | Peptide conc. | Proteinase act. |
ACEI act. | Change in SBP | ||
(%) | (U/ml) | (U/ml) | (mmHg) | |||
Non-fermented milk | 0.00 | - | 0 | - 5.0 ± 7.3 | ||
(Lactobacilli) | ||||||
L. helveticusCP790 | 0.19 | 230 | 58 | - 27.4 ± 13.3 ** | ||
L. helveticusCP611 | 0.25 | 367 | 70 | - 20.0 ± 9.6 ** | ||
L. helveticusCP615 | 0.18 | 420 | 51 | - 23.0 ± 13.4 ** | ||
L. helveticus JCM1006 | 0.15 | 182 | 26 | - 15.2 ± 9.3 * | ||
L. helveticusJCM1120 | 0.10 | 112 | 34 | - 6.5± 10.8 | ||
L. helveticusJCM1004 | 0.21 | 186 | 48 | - 29.3 ± 13.6 ** | ||
L. delbrueckii subsp.bulgaricus CP973 | 0.19 | 105 | 22 | - 0.8 ± 8.2 | ||
L. delbrueckii subsp.bulgaricus JCM1002 | 0.11 | 124 | 28 | - 4.5 ± 4.0 | ||
L. casei CP680 | 0.01 | 35 | 3 | - 0.2 ± 6.6 | ||
L. casei JCM1134 | 0.00 | 28 | 9 | - 7.0 ± 11.2 | ||
L. casei JCM1136 | 0.09 | 25 | 18 | - 9.6 ± 7.2 | ||
L. acidophilus JCM1132 | 0.00 | 28 | 8 | - 8.7 ± 7.8 | ||
L. delbrueckii subsp. lactisJCM1105 | 0.08 | 18 | 16 | - 3.3 ± 3.5 | ||
(Streptococci) | ||||||
S. thermophilus CP1007 | 0.02 | 25 | 3 | - 2.4 ± 8.1 | ||
(Lactococci) | ||||||
L. lactis subsp. lactis CP684 | 0.00 | 35 | 4 | - 7.3 ± 10.5 | ||
L. lactis subsp. cremoris CP312 | 0.02 | 18 | 4 | - 5.8 ± 13.9 | ||
Significant differences from the control, p< 0.01, p< 0.05. | ||||||
ACEI activity: Peptides that show 50% inhibition of ACE activity was defined as one unit. | ||||||
SBP: systolic blood pressure of spontaneously hypertensive rat |
1.2. Clinical effects of the fermented milk
Hypertension is a major risk factor in cardiovascular diseases, such as heart disease and stroke. In order to reduce the incidence of disease, pharmacological substances can be used to decrease high blood pressure to within the normal range. In the first Japanese study with the fermented milk, hypertensive subjects were randomly assigned to two groups: the one group ingested 95 ml of the milk, containing 3.4 mg of VPP and IPP, daily for 8 wk; the other group ingested the same amount of artificially acidified milk as a placebo, for 8 wk (30). In the fermented milk group, systolic blood pressure decreased significantly between 4 and 8 wk after the beginning of ingestion, but not in the placebo group (30). Moreover, clinical tests were performed for Japanese subjects with different blood pressure levels, which confirmed the mild and prolonged effects for the hypertensive subjects following oral administration of bioactive milk(30-33) (Fig. 1). In a pilot study conducted in Finland, the antihypertensive effect was also observed in the group ingesting
1.3. Processing of antihypertensive peptides in L. helveticus
1.3.1. Proteolytic system in lactic acid bacteria
Many kinds of proteolytic enzymes have been reported from lactic acid bacteria, and were reviewed extensively(26, 27, 36, 37). The components of the proteolytic systems of lactic acid bacteria are divided into three groups, including the extracellular proteinase that catalyzes casein breakdown to peptides, peptidases that hydrolyze peptides to amino acids and a peptide transport system. The number of proteinases was reported from lactococci, which are mainly used in cheese making. The extracellular proteinase activity is linked to cell growth in milk and seemed to be essential for utilization of milk protein for growth. The gene encoding the proteinases, named
1.4. Processing by a cell wall-associated proteinase
The first step in casein decomposition is typically caused by an extracellular proteinase, and further digestion to amino acids is catalyzed by many kinds of intracellular peptidases (25, 36). Among the lactic acid bacteria,
Moreover, a slight difference in the specificity of the two types of proteinases toward casein was suggested for the two types of
No | Strain | Subspecies | Reactivity (CP790)1 | Reactivity (CP53)2 | Type | |
1 | L. helveticus CP39 | J3 | 45 kDa | 45 kDa | A | |
2 | L. helveticus CP53 | H4 | ND5 | 53, 170 kDa | B | |
3 | L. helveticus CP209 | J | 45 kDa | 45 kDa | A | |
4 | L. helveticus CP210 | J | 45 kDa | 45 kDa | A | |
5 | L. helveticus CP293 | J | 45 kDa | 45 kDa | A | |
6 | L. helveticus CP510 | J | 45 kDa | 45 kDa | A | |
7 | L. helveticus CP611 | J | 45 kDa | 45 kDa | A | |
8 | L. helveticus CP615 | J | 45 kDa | 45 kDa | A | |
9 | L. helveticus CP617 | J | 45 kDa | 45 kDa | A | |
10 | L. helveticus CP789 | J | 45 kDa | 45 kDa | A | |
11 | L. helveticus CP790 | J | 45 kDa | 45 kDa | A | |
12 | L. helveticus JCM1004 | H | ND | 53, 170 kDa | B | |
13 | L. helveticus JCM1006 | J | 45 kDa | 45 kDa | A | |
14 | L. helveticus JCM1007 | J | ND | 53, 170 kDa | B | |
15 | L. helveticus JCM1062 | J | ND | 53, 170 kDa | B | |
16 | L. helveticus JCM1103 | H | ND | 53, 170 kDa | B | |
17 | L. helveticus JCM1120 | H | ND | 53, 170 kDa | B | |
(Yamamoto et al., 1998, Biosci.Biotech.Biochem. 58, 776-778) | ||||||
1Monoclonal antibody to the proteinase from L. helveticus CP790 | ||||||
2Polyclonal antibody to the proteinase from L. helveticus CP53 | ||||||
3Classified as L. helveticusbiovarjugurti | ||||||
4Classified as L. helveticusbiovarhelveticus | ||||||
5Not detected |
Currently, we completed the whole genome sequence of
Proteolytic enzyme | Gene | Molecular weight (kDa) | Protein ID | Identity(%) | |
CM4 | DPC4571 | DPC4571 | |||
Proteinase | prtY | 47.0 | ND | - | - |
prtH2 | 181.6 | 180.871 | - | 99.2 | |
prtM | 33.7 | 32.7 | ABX27563 | 98.0 | |
Aminopeptidase | pepC1 | 51.4 | 51.4 | ABX26582 | 98.9 |
pepC2 | 52.9 | 50.2 | ABX27065 | 98.6 | |
pepN | 95.8 | 95.9 | ABX27731 | 99.4 | |
pepN2 | 57.2 | 57.2 | ABX27544 | 100.0 | |
pepA | 41.3 | 40.1 | ABX26758 | 99.4 | |
XPDAP | pepX | 90.5 | 90.6 | ABX27419 | 99.6 |
Endopeptidase | pepE | 50.0 | 50.0 | ABX26466 | 99.8 |
pepE2 | 51.4 | 50.3 | ABX26457 | 99.8 | |
pepF | 68.1 | 68.1 | ABX27686 | 99.3 | |
pepO | 73.6 | 73.5 | ABX27358 | 99.4 | |
pepO2 | 73.8 | 73.5 | ABX27211 | 98.6 | |
pepO3 | 73.1 | 72.6 | ABX26433 | 99.7 | |
Tripeptidase | pepT | 47.1 | 46.7 | ABX27305 | 99.3 |
pepT2 | 48.8 | 48.4 | ABX27165 | 99.5 | |
Dipeptidase | pepD1 | 54.0 | 54.1 | ABX27625 | 99.4 |
pepD2 | 54.9 | 54.9 | ABX27375 | 99.8 | |
pepD3 | 53.5 | 53.5 | ABX27723 | 100.0 | |
pepV | 51.5 | 51.5 | ABX27224 | 98.9 | |
pepDA | 53.5 | 53.5 | ABX26492 | 99.6 | |
Prolidase | pepQ | 41.2 | 41.2 | ABX26664 | 99.5 |
pepQ2 | 41.4 | 41.1 | ABX27405 | 99.7 | |
Prolinase | pepPN | 35.0 | 35.0 | ABX27633 | 99.7 |
Prolineiminopeptidase | pepI | 33.9 | 33.8 | ABX26375 | 99.3 |
ND: Not detected, *size of the reported pseudo-gene |
By the proteolytic action of the extracellular proteinase in CP790 (and CM4), a long -casein peptide with a 28 amino acid residue including VPP and IPP sequences was generated (29) (Fig. 2). The proteinase activity is easily repressed by accumulated peptides by the proteinase in the fermented milk. Moreover, the enzyme activity is inactivated by pH drop during the fermentation. So, the first degradation of casein by the extracellular proteinase would be occurred mostly at the beginning of the fermentation.
1.4.1. Intracellular processing by some peptidases
Next, the long peptide was thoroughly hydrolyzed to shorter peptides by intracellular peptidases. Intracellular peptidases of
Based on the previous reported characteristics of many peptidases from
1.5. Comparison of the L. helveticus proteolytic system to those in other lactic acid bacteria
The unprocessed proteinase of
1.6. Repression of proteolyticsystemg
For growth of lactic acid bacteria in milk, the proteolytic system is activated in the milk medium because of alimited amount of amino acids. However, during fermentation in the milk medium,the proteolytic system of lactic acid bacteria is repressed by accumulated peptides in the fermented milk. The amount of VPP and IPP in the
2. Conclusion
In this paper, we showed the potential of bioactive peptides to maintain blood pressure in the normal range. About 30% of Japanese people are estimated to be at risk for hypertension. Generally, hypertension has been improved by medication and partly by controlling the diet. Recently, some food products containing antihypertensive peptides and proven antihypertensive effects in clinical studies were recognized as functional foods, Foods for Specified Health Use (FOSHU) in Japan. Biologically functional peptides exerting a mild influence on hypertensive subjects without adverse effect have enormous potential in reducing the risk of cardiovascular disease.
Among many kinds of commercially available lactic acid bacteria,
References
- 1.
Migliore-Samour D. Floćh F. Jollès P. 1989 Biologically active casein peptides implicated in immunomodulation. J. Dairy Res., 56, 357-362 (1989). - 2.
Bellamy W. Takase M. Yamauchi K. Shimamura S. Tomita M. 1992 Identification of the bactericidal domain of lactoferrin. ,1121,130 6 (1992). - 3.
Teschemacher H. Koch G. Brantl V. Milk protein-derived opioid receptor ligands. , 43,99 117 (1997). - 4.
FitzGerald R. J. 1998 Potential uses of caseinophosphopeptides . , 8,451 457 (1998). - 5.
Yamamoto N. 1997 Antihypertensive peptides derived from food proteins. , 43,129 34 (1997). - 6.
Meisel H. Bockelmann W. 1999 Bioactive peptides encrypted in milk proteins: proteolytic activation and thropho-functional properties. , 76,207 15 (1999). - 7.
Yamamoto N. Takano T. 1999 Antihypertensive peptides derived from milk proteins. Nahrung, 43,159 64 (1999). - 8.
Takano T. 2002 Anti-hypertensive activity of fermented dairy products containing biogenic peptides. , , 82,333 40 (2002). - 9.
Caldwell P. R. Seegal B. C. Hsu K. C. Das M. Soffer R. L. 1976 Angiotensin-converting enzyme: vascular endothelial localization. Science, 191, 1050-1051 (1976). - 10.
El-Dorry H. A. Bull H. G. Iwata K. Thornberry N. A. Cordes E. H. Sofffer R. L. 1982 Molecular and catalytic properties of rabbit testicular dipeptidyl carboxypeptidase., J. Biol. Chem.,257, 14128-14133 (1982). - 11.
Skeggs L. T. J. Kahn J. R. Shumway N. P. 1956 The preparation and function of the hypertensin-converting enzyme. J. Exp. Med., 103,295 9 (1956). - 12.
Yang H. Y. T. Erdos E. G. Levin Y. 1970 A dipeptidyl carboxypeptidase that converts angiotensin I and inactivates bradykinin. Biochimica et Biophysica Acta, 214, 374-376 (1970). - 13.
Ferreira S. H. Bartelt D. C. Greene L. J. 1970 Isolation of bradykinin-potentiating peptides from Bothrops jararaca venom. , 9, 2583-2259 (1970). - 14.
Ondetti M. A. Williams N. J. Sabo E. F. Plušcec J. Weaver E. R. Kocy O. 1971 Angiotensin-converting enzyme inhibitors from the venom of Bothrops jararaca. Isolation, elucidation of structure, and synthesis. , 10, 4033-4039 (1971). - 15.
Yamamoto N. Akino A. Takano T. 1994 Antihypertensive effect of different kinds of fermented milk in spontaneously hypertensive rats. Biosci. Biotech. Biochem., 58, 776-778 (1994). - 16.
Yamamoto N. Akino A. Takano T. 1994 Antihypertensive effect of the peptides derived from casein by an extracellular proteinase from Lactobacillus helveticus CP790. , 77, 917-922 (1994). - 17.
Maruyama S. Mitachi H. Tanaka H. Tomizuka N. Suzuki H. 1987 Study on the active site and antihypertensive activity of angiotensin I-converting enzyme inhibitors derived from casein. Agric. Biol. Chem., 51, 1581-1586 (1987). - 18.
Kohmura M. Nio N. Kubo K. Minoshima Y. Nunekata E. Ariyoshi Y. 1989 Inhibition of angiotensin-converting enzymes by synthetic peptides of human beta-casein. Agric. Biol. Chem., 53, 2107-2114 (1989). - 19.
Nakamura Y. Yamamoto N. Sakai K. Okubo A. Yamazaki S. Takano T. 1995 Purification and characterization of angiotensin I-converting enzyme inhibitors from sour milk. J. Dairy Sci., 78, 777-783 (1995). - 20.
Yamamoto N. Maeno M. Takano T. 1999 Purification and characterization of an antihypertensive peptide from a yogurt-like product fermented by CPN4. J. Dairy Sci., 82, 1388-1393 (1999). - 21.
Maeno M. Yamamoto N. Takano T. 1996 Identification of an antihypertensive peptide from casein hydrolysate produced by a proteinase from Lactobacillus helveticus CP790. , 79, 1316-1321 (1996). - 22.
Abubakar A. Saito T. Kitazawa H. Kawai Y. Itoh T. 1998 Structural analysis of new antihypertensive peptides derived from cheese whey protein by proteinase K digestion. J. Dairy Sci., 81, 3131-3138 (1998). - 23.
Pihlanto-Leppala A. Koskinen P. Piilola K. Tupasela T. Korhonen H. 2000 Angiotensin I-converting enzyme inhibitory properties of whey protein digests: concentration and characterization of active peptides. J. Dairy Res., 67, 53-64 (2000). - 24.
Murakami M. Tonouchi H. Takahashi R. Kitazawa H. Kawai Y. Negishi H. 2004 Saito, T. Structural analysis of a new anti-hypertensive peptide (beta-lactosin B) isolated from a commercial whey product. J. Dairy Sci., 87, 1967-1974 (2004). - 25.
Smid E. J. Poolman B. Konings W. N. 1991 Casein utilization by lactococci. Appl. Environ. Microbiol., 57, 2447-2452 (1991). - 26.
Pritchard G. G. Coolbear T. 1993 The physiology and biochemistry of the proteolytic system in lactic acid bacteria. FEMS Microbiol. Rev., 12, 179-206 (1993). - 27.
Tan P. S. Poolman B. Konings W. N. 1993 Proteolytic enzymes of Lactococcus lactis. , 60, 269-286 (1993). - 28.
Nakamura Y. Yamamoto N. Sakai K. Takano T. 1995 Antihypertensive effect of sour milk and peptides isolated from it that are inhibitors to angiotensin I-converting enzyme. J. Dairy Sci., 78, 1253-1257 (1995). - 29.
Yamamoto N. Akino A. Takano T. 1993 Purification and specificity of a cell-wall-associated proteinase from Lactobacillus helveticus CP790. , 114, 740-745 (1993). - 30.
Hata Y. Yamamoto M. Ohni M. Nakajima K. Nakamura Y. Takano T. 1996 A placebo-controlled study of the effect of sour milk on blood pressure in hypertensive subjects. Am. , 64, 767-771 (1996). - 31.
Kajimoto O. Aihara K. Hirata H. Takahashi R. Nakamura Y. 2001 Hypotensive Effects of Tablets Containing "lactotripeptides (VPP, IPP)". J. Nutr. Food (in Japanese), 4, 51-61 (2001). - 32.
Kajimoto O. Aihara K. Hirata H. Takahashi R. Nakamura Y. 2001 Safety evaluation of excessive intake of the tablet containing "lactotripeptides (VPP, IPP)" in healthy volunteers. J. Nutr. Food (in Japanese), 4, 37-46 (2001). - 33.
Nakamura Y. Kajimoto O. Aihara K. Mizutani J. Ikeda N. Nishimura A. Kajimoto Y. 2004 Effects of the liquid yogurts containing "lactotripeptide (VPP,IPP)" on high-normal blood pressure. J. Nutr. Food (in Japanese), 7, 123-137 (2004). - 34.
Seppo L. Jauhiainen T. Poussa T. Korpela R. 2003 A fermented milk high in bioactive peptides has a blood pressure-lowering effect in hypertensive subjects. Am. J. Clin. Nutr., 77, 326-330 (2003). - 35.
Seppo L. Kerojoki O. Suomalainen T. Korpela R. 2002 The effect of a Lactobacillus helveticus LBK-16 H fermented milk on hypertension- a pilot study on humans. , 57, 124-127 (2002). - 36.
Kunji E. R. Mierau I. Hagting A. Poolman B. Konings W. N. 1996 The proteolytic systems of lactic acid bacteria. Antonie Van Leeuwenhoek, 70, 187-221 (1996). - 37.
Christensen J. E. Dudley E. G. Pederson J. A. Steele J. L. 1999 Peptidases and amino acid catabolism in lactic acid bacteria. Antonie Van Leeuwenhoek, 76, 217-246 (1999). - 38.
Kok J. Venema G. 1988 Genetics of proteinases of lactic acid bacteria. Biochimie, 70, 475-488 (1988). - 39.
Kiwaki M. Ikemura H. Shimizu-Kadota M. Hirashima A. 1989 Molecular characterization of a cell wall-associated proteinase gene from Streptococcus lactis NCDO763. ., 3, 359-369 (1989). - 40.
de Vos W. M. Vos P. de Haard H. Boerrigter I. 1989 Cloning and expression of the Lactococcus lactis subsp. cremoris SK11 gene encoding an extracellular serine proteinase. , 85, 169-176 (1989). - 41.
Yamamoto N. Ono H. Maeno M. Takano T. 1998 Classification of Lactobacillus helveticus strains by Immunological differences in extracellular proteinases. , 62, 1228-1230 (1998). - 42.
Yamamoto N. Akino A. Takano T. Shishido K. 1995 Presence of active and inactive molecules of a cell wall-associated proteinase in Lactobacillus helveticus CP790. , 61, 698-701 (1995). - 43.
Pederson J. A. Mileski G. J. Weimer B. C. Steele J. L. 1999 Genetic characterization of a cell envelope-associated proteinase from Lactobacillus helveticus CNRZ32. , 181, 4592-4597 (1999). - 44.
Yamamoto N. Shinoda T. Takano T. 1999 Molecular cloning and sequence analysis of a gene encoding an extracellular proteinase from Lactobacillus helveticus CP790. , 64, 1217-1222 (1999). - 45.
Yamamoto N. Takano T. 1997 Maturation protein need for activation of an extracellular proteinase in Lactobacillus helveticus CP790. , 80, 1949-1954 (1997). - 46.
Ono H. Yamamoto N. Maeno M. Takano T. 1997 Purification and characterization of a cell-wall associated proteinase from Lactobacillus helveticus CP53. , 52, 373-377 (1997). - 47.
Yamamoto N. Ishida Y. Kawakami N. Yada H. 1991 Lactobacillus helveticus bacterium having high capability of producing tripeptide, fermented milk product, and process for preparing the same. EU Patent, 1016709A1 (1991). - 48.
Callanan M. Kaleta P. O’Callaghan J. O’Sullivan O. Jordan K. McAuliffe O. Sangrador-Vegas A. Slattery L. Fitzgerald G. F. Beresford T. Ross R. P. 2008 Genome sequence of Lactobacillus helveticus, an organism distinguished by selective gene loss and insertion sequence element expansion. 190:727-735 (2008). - 49.
Shinoda T. Wakai T. Uchida Hattori M. Nakamura Y. Yamamoto N. 2011 Comparative Analysis of Proteolytic Enzymes Need for Processing of Antihypertensive Peptides between Lactobacillus helveticus CM4 and DPC4571. In preparatoin for the submission to publicatoin (2011). - 50.
Genay M. Sadat L. Gagnaire V. Lortal S. 2009 prtH2, not prtH, is the ubiquitous cell wall proteinase gene in Lactobacillus helveticus. 75:3238-3249 (2009). - 51.
Pederson J. A. Mileski G. J. Weimer B. C. Steele J. L. 1999 Genetic characterization of a cell envelope-associated proteinase from Lactobacillus helveticus CNRZ32. 181: 4592-4597 (1999). - 52.
Ueno K. Mizuno S. Yamamoto N. 2004 Purification and characterization of an endopeptidase has an important role in the carboxyl terminal processing of antihypertensive peptides in Lactobacillus helveticus CM4. , 39, 313-318 (2004). - 53.
Yamamoto N. Shinoda T. Mizuno S. 2004 Cloning and expression of an endopeptidase gene from Lactobacillus helveticus CM4 involved in processing antihypertensive peptides. , 59, 593-597 (2004). - 54.
Altermann E. Russell W. M. Azcarate-Peril M. A. Barrangou R. Buck B. L. McAuliffe O. Souther N. Dobson A. Duong T. Callanan M. Lick S. Hamrick A. Cano R. Klaenhammer T. R. 2005 Complete genome sequence of the probiotic lactic acid bacterium Lactobacillus acidophilus NCFM. , 102, 3906-3912 (2005). - 55.
DOE Joint Genome Institute 2002 Lactobacillus gasseri whole genome shotgun sequencing project. (2002). - 56.
Pridmore R. D. Berger B. Desiere F. Vilanova D. Barretto C. Pittet A. C. Zwahlen M. C. Rouvet M. Altermann E. Barrangou R. Mollet B. Mercenier A. Klaenhammer T. Arigoni F. Schell M. A. 2004 The genome sequenc e of the probiotic intestinal bacterium Lactobacillus johnsonii NCC 533. , 101, 2512-2517 (2004). - 57.
Wakai T. Yamamoto N. - 58.
den Hengst C. D. Curley P. Larsen R. Buist G. Nauta A. van Sinderen D. Kuipers O. P. Kok J. 2005 Probing direct interactions between CodY and the oppD promoter of Lactococcuslactis. 187: 512-521 (2005). - 59.
den Hengst C. D. van Hijum S. A. Geurts J. M. Nauta A. Kok J. Kuipers O. P. 2005 The LactococcuslactisCodYregulon: identification of a conserved cis-regulatory element. 280: 34332-34342 (2005).