Optimization of nisin concentration in fermented beef sausage to decrease sodium nitrite concentration in the final product
Subject Areas : Food MicrobiologyFatemeh Vaheb 1 , Nafiseh Sadat Naghavi 2 , Zarrindokht Emami-Karvani 3
1 - Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran
2 - Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran
3 - Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran
Keywords: Microbial count, Nisin, Nitrite, Fermented beef,
Abstract :
Bacteriocins such as nisin produced by lactic acid bacteria exhibit antimicrobial activity at very low concentrations without side effects. The aim of the present study was to optimize nisin content in fermented beef to achieve optimum pH and microbial community as well as reduction of nitrite concentration in the final product. In this experimental laboratory research, fermented beef was prepared using Lactobacillus plantarum and Lactobacillus sakei. Then, optimization of nisin and sodium nitrite concentrations were done by response surface methodology based on the pH decrease and spoilage causing microbial count in fermented product. Among treatments, 3 treatments with respectively 80 and 100 ppm sodium nistrit and nisin, 128.28 and 140 ppm sodium nitrite and nisin and 180 and 196 ppm sodium nistrit and nisin showed the best results based on pH and spoilage causing microbial count reduction. The residual nitrite contents in these three treatments reduced to 5.7 to 8.47 ppm, which were lower than the limited amount. Clostridium was absent in all three beef products after one mount storage at 4 °C. Addition of nisin to fermented beef significantly decreased the nitrite concentration. Therefore, the usage of nisin is recommended as a preservative in beef sausage industries in Iran.
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2. Ferreira IMPL, Silva S. uantification of residual nitrite and nitrate in ham by reverse phase
high performance liquid choromatography/diode detector. Talanta. 2018; 74: 1598-1602.
3. Aran H , Biscola , Jaffrès E Dousset , Pillot G. Bacteriocin-producing nterococcus
aecalis T2W2G isolated from mangrove forests in southern Thailand: purification
characterization and safety evaluation. Food Control. 2015; 54: 126-134.
4. Birri DJ, Brede DA, Forberg T, Holo H, Nes IF. Molecular and genetic characterization of a
novel bacteriocin locus in nterococcus avium isolates from infants. Appl Environ Microbiol.
2010; 76: 483-492.
5. Solomakos N, Govaris A, oidis P, Botsoglou N. The antimicrobial effect of thyme essential
oil, nisin and their combination against sc eric ia coli O157:H7 in minced beef during
refrigerated storage. Meat Sci. 2008; 80: 159-166.
6. Monica F, Sara C. Yarrowia lipolytica strains in dried fermented sausage manufacture. J Meat
Sci. 2015; 75: 669-675.
7. Sadler GD, Murphy PA. pH and titratable acidity. In: Nielsen SS, editors. Food analysis. 3rd ed.
New York. Springer Science and Business Media Publishers; 2003: 207–225.
8. Alvarez-Sieiro P, Montalbán-López M, Mu D, uipers OP. Bacteriocins of lactic acid bacteria:
extending the family. Appl Microbiol Biotechnol. 2016; 100(7): 2939-51.
9. Hereu A, Bover-Cid S, Garriga M, Aymerich T. High hydrostatic pressure and biopreservation
of dry-cured ham to meet the Food Safety Objectives for Listeria monocyto enes. Int J Food
Microbiol. 2012; 154 (3): 107-112.
10. ahnama E, Naghavi NS. Optimization of fermented cow meat quality by lactic acid bacteria
in batch fermentation JMW. 2017; 10(3): 263-274.
11. Wang W, Wang H, en HY, Liu DY, Zhu WY. Effect of inoculating Lactobacillus sakei
starter cultures on themicrobiological quality and nitrite depletion of Chinese fermented
sausages. J Food Control. 2013; 32: 591-596.
12. hairul Anwar M S, Mohamed Afizal M A. Overview on the response surface methodology
( SM) in extraction processes. J. Appl. Sci. Eng. 2015; 2(1): 1-10.
13. iazi F, Zeynali F, Hoseini H, Behmadi H. Effect of dry red grape pomace as a nitrite
substitute on the microbiological and physicochemical properties and residual nitrite of
dry-cured sausage. Nut Food Sci es. 2016; 3(3): 37-44.
14. Jansen S, Dera T, uth MS, Syofyasti MN, Yosy CS. Analysis of nitrite and nitrate in the
corned beef and smoked beef by using visible spectrophotometry method. InIOP Conference
Series: Earth and Environ Sci. 2018; 205(1): 012039.
15. Harris LJ, Fleming HP, laenhammer T . Developments in nisin research. Food es Int.
1992; 25: 57-66.
16. Pajohi M , Tajik H, Farshid AA. Antimicrobial activity of Nisin on acillus cereus and
acillus subtilis in a food model and their ultra structural investigation. Iranian J Med
Microbiol. 2010; 4(3): 45-52. [n Persian]
17. indhe SN, Shinde AT, Jadhav BA, umbhar H. Application of hurdle technology for
preservation of chiken susage at ambient temperature (37±1 °C). Int J Pure Appl Biosci. 2017;
5(4): 480-488.
18. ara , Yaman H, Gök , Akkaya L. The effect of nisin on Listeria monocyto enes in
chicken burgers. Indian J Anim es. 2014; 48 (2): 171-176.
19. Hampikyan H, Ugur M. The effect of nisin on L. monocyto enes in Turkish fermented
sausages (sucuks). Meat Sci. 2007; 76(2): 327-332.
20. ayman M , Aris B, Hurst A. Nisin: a possible alternative or adjunct to nitrite in the
preservation of meats. Appl Environ Microbiol. 1981; 41(2): 375-380.
21. iong Y, Chen M, Warner D, Fang Z. Incorporating nisin and grape seed extract in
chitosan-gelatine edible coating and its effect on cold storage of fresh pork. Food Control.
2020; 110: 107018.
22. Churklam W, Chaturongakul S, Ngamwongsatit B, Aunpad . The mechanisms of action of
carvacrol and its synergism with nisin against Listeria monocyto enes on sliced bologna
sausage. Food Control. 2020; 108: 106864.
23. Nguyen T, Brody H, Lin GH, angé H, uraji , Ye C, amarajan P, adaic A, Gao L,
apila Y. Probiotics, including nisin‐based probiotics, improve clinical and microbial outcomes
relevant to oral and systemic diseases. Periodontol. 2020; 82(1): 173-85.
24. Hammes WP. Metabolism of nitrate in fermented meats: the characteristic feature of a specific
group of fermented foods. Food Microbiol. 2012; 29(2): 151-6.
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nitrite/nitratein foods: NM L collaborative study. J AOAC Int. 2010; 83 (2): 365-75.
2. Ferreira IMPL, Silva S. uantification of residual nitrite and nitrate in ham by reverse phase
high performance liquid choromatography/diode detector. Talanta. 2018; 74: 1598-1602.
3. Aran H , Biscola , Jaffrès E Dousset , Pillot G. Bacteriocin-producing nterococcus
aecalis T2W2G isolated from mangrove forests in southern Thailand: purification
characterization and safety evaluation. Food Control. 2015; 54: 126-134.
4. Birri DJ, Brede DA, Forberg T, Holo H, Nes IF. Molecular and genetic characterization of a
novel bacteriocin locus in nterococcus avium isolates from infants. Appl Environ Microbiol.
2010; 76: 483-492.
5. Solomakos N, Govaris A, oidis P, Botsoglou N. The antimicrobial effect of thyme essential
oil, nisin and their combination against sc eric ia coli O157:H7 in minced beef during
refrigerated storage. Meat Sci. 2008; 80: 159-166.
6. Monica F, Sara C. Yarrowia lipolytica strains in dried fermented sausage manufacture. J Meat
Sci. 2015; 75: 669-675.
7. Sadler GD, Murphy PA. pH and titratable acidity. In: Nielsen SS, editors. Food analysis. 3rd ed.
New York. Springer Science and Business Media Publishers; 2003: 207–225.
8. Alvarez-Sieiro P, Montalbán-López M, Mu D, uipers OP. Bacteriocins of lactic acid bacteria:
extending the family. Appl Microbiol Biotechnol. 2016; 100(7): 2939-51.
9. Hereu A, Bover-Cid S, Garriga M, Aymerich T. High hydrostatic pressure and biopreservation
of dry-cured ham to meet the Food Safety Objectives for Listeria monocyto enes. Int J Food
Microbiol. 2012; 154 (3): 107-112.
10. ahnama E, Naghavi NS. Optimization of fermented cow meat quality by lactic acid bacteria
in batch fermentation JMW. 2017; 10(3): 263-274.
11. Wang W, Wang H, en HY, Liu DY, Zhu WY. Effect of inoculating Lactobacillus sakei
starter cultures on themicrobiological quality and nitrite depletion of Chinese fermented
sausages. J Food Control. 2013; 32: 591-596.
12. hairul Anwar M S, Mohamed Afizal M A. Overview on the response surface methodology
( SM) in extraction processes. J. Appl. Sci. Eng. 2015; 2(1): 1-10.
13. iazi F, Zeynali F, Hoseini H, Behmadi H. Effect of dry red grape pomace as a nitrite
substitute on the microbiological and physicochemical properties and residual nitrite of
dry-cured sausage. Nut Food Sci es. 2016; 3(3): 37-44.
14. Jansen S, Dera T, uth MS, Syofyasti MN, Yosy CS. Analysis of nitrite and nitrate in the
corned beef and smoked beef by using visible spectrophotometry method. InIOP Conference
Series: Earth and Environ Sci. 2018; 205(1): 012039.
15. Harris LJ, Fleming HP, laenhammer T . Developments in nisin research. Food es Int.
1992; 25: 57-66.
16. Pajohi M , Tajik H, Farshid AA. Antimicrobial activity of Nisin on acillus cereus and
acillus subtilis in a food model and their ultra structural investigation. Iranian J Med
Microbiol. 2010; 4(3): 45-52. [n Persian]
17. indhe SN, Shinde AT, Jadhav BA, umbhar H. Application of hurdle technology for
preservation of chiken susage at ambient temperature (37±1 °C). Int J Pure Appl Biosci. 2017;
5(4): 480-488.
18. ara , Yaman H, Gök , Akkaya L. The effect of nisin on Listeria monocyto enes in
chicken burgers. Indian J Anim es. 2014; 48 (2): 171-176.
19. Hampikyan H, Ugur M. The effect of nisin on L. monocyto enes in Turkish fermented
sausages (sucuks). Meat Sci. 2007; 76(2): 327-332.
20. ayman M , Aris B, Hurst A. Nisin: a possible alternative or adjunct to nitrite in the
preservation of meats. Appl Environ Microbiol. 1981; 41(2): 375-380.
21. iong Y, Chen M, Warner D, Fang Z. Incorporating nisin and grape seed extract in
chitosan-gelatine edible coating and its effect on cold storage of fresh pork. Food Control.
2020; 110: 107018.
22. Churklam W, Chaturongakul S, Ngamwongsatit B, Aunpad . The mechanisms of action of
carvacrol and its synergism with nisin against Listeria monocyto enes on sliced bologna
sausage. Food Control. 2020; 108: 106864.
23. Nguyen T, Brody H, Lin GH, angé H, uraji , Ye C, amarajan P, adaic A, Gao L,
apila Y. Probiotics, including nisin‐based probiotics, improve clinical and microbial outcomes
relevant to oral and systemic diseases. Periodontol. 2020; 82(1): 173-85.
24. Hammes WP. Metabolism of nitrate in fermented meats: the characteristic feature of a specific
group of fermented foods. Food Microbiol. 2012; 29(2): 151-6.
