Protein Quality of Fermented Beef by Lactobacillus plantarum 1B1

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575 Proceeding of the 2nd_{International Seminar on Animal Industry | Jakarta, 5-6 July 2012}

Protein Quality of Fermented Beef by

Lactobacillus plantarum 1B1

I. I. Arief*, R.R.A. Maheswari, T. Suryati, & N. Kurniawati

Department of Animal Production and Technology, Faculty of Animal Science, Bogor Agricultural University, Bogor, 16680, Indonesia

*e-mail: irma_isnafia@yahoo.com

Abstract

This study was conducted to investigate the proteolytic activity and protein characteristics of fermented beef. The effect of different mechanical treatments on beef, (sliced and ground beef) was compared. The contents of amino acid were analyzed using HPLC (high performance liquid chromatography) method. The results indicated that Lactobacillus plantarum 1B1, isolated from beef, had better proteolytic activity on sliced fermented beef if compared to ground fermented beef, and improved crude protein on sliced fermented beef. Sliced and groud fermented beef contained 15 components of amino acids which consisted of threonine, tyrosine, methionine, valine, phenylalanine, isoleucine, leucine and lysine as essential amino acid and aspartate, glutamate, serine, histidine, glycine, alanine and arginine as non-essential amino acids. Leucine was the highest among essential amino acids content and glutamate was the highest among non-essential amino acids content. The SDS PAGE profiles showed the beef protein molecule weight range of 94.71 - 13.43 kD. The numbers of protein band of fresh beef, sliced and ground fermented beef were 12, 7 and 7. In conclusion, fermentation process on raw beef by Lactobacillus plantarum could improve amino acid availability because of its proteolytic activity on beef protein.

Key words: fermented beef, Lactobacillus plantarum, protein quality

Introducton

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process by lactc acd bactera, such as Lactobacillus plantarum (L. plantarum). The fermentaton mght extend the shelf lfe and appearance of meat products. The addton of starter L. plantarum 1B1 n beef fermentaton s expected to perform on the proteolytc actvty of beef. The am of ths research was to nvestgate the qualty of proten of fermented beef by L. plantarum 1B1.

Materals and Methods

L. plantarum 1B1 was obtaned from Laboratory of anmal product and technology, Faculty of Anmal Scence, Bogor Agrcultural Unversty. The stran was solated from local beef and was dentfed usng 16S rRNA gene sequencng (Aref, 2011).

The treatment used n ths experment was type of beef fermentaton. DFD beef (pH value : 6.2) was used n ths experment. Slced and grnded beef wth no-culated by L. plantarum 1B1 for fermentaton. Starter culture of 10% w/w L. plan-tarum 1B1 were noculated nto fermentaton process. The slced beef was arranged n a bakng dsh coated wth alumnum fol and covered wth plastc top surface. The ground beef was put n a plastc bag, flaked wth a thckness of approxmately 3 mm, packed and arranged n a bakng dsh. These treated meat groups were then ncubated for 24 h at 37°C, smoked for nne hours at a temperature of 30o_C.

Varables observed ncluded the content of crude proten (AOAC, 2005), amno acd composton usng HPLC (AOAC, 2005), proteolytc actvty of L. plantarum

1B1 (Bergmeyer and Gawehn (1983), and SDS-PAGE electrophoress of protenBergmeyer and Gawehn (1983), and SDS-PAGE electrophoress of proten, and SDS-PAGE electrophoress of proten from fermented beef (Fadda et al., 1998).

The experment was set up n a completely randomzed desgn wth three replcaton. The data were analyzed usng ANOVA, and f found any dfferences, they were further tested usng Tukey test (Steel and Tore, 1995). Amno acd composton and SDS Page electrophoress were descrbed by descrptve analyss.

Results and Dscusson

Crude protein of fermented beef

Protens n food generally determne the qualty of a product prmarly derved from meat. Isolates of L. plantarum 1B1 used to acheve optmzaton of beef fermentaton wth the assumpton that these bactera would be more adaptve to beef. Crude proten and proteolytc actvty of of fermented beef s presented n Table 1.

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that of ground fermented beef. Grndng lossed sarcoplasmc proten of beef t could reduce total proten content of ground beef.

Proteolytc actvty of fresh beef was not detected (0), t means that natural proteolytc enzyme of beef was not actve. Increased proteolytc actvty occurred after the fermentaton took place n fermented beef. Ths proved that the solates of L. plantarum 1B1 had proteolytc actvty on beef proten. L. plantarum 1B1 had extracelullar protease enzyme n the de_man Rogosa Sharp broth 0.041 U/ml. Accordng to Fadda et al. (1998) that some strans of Lactobacillus bactera such as

L. casei and L. plantarum can produce proteolytc enzymes n fermented meats.

Molecular weight protein of fermented beef

Fermentaton could degrade beef proten, caused by proteolytc actvty of L. plantarum 1B1, as shown n Fgure 1.

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Fgure 1. SDS-PAGE electrophoress bands of beef proten (n= marker; A= fresh DFD beef; B=slced fermented beef; C=grnded fermented beef).

Ths type of proten bands detected n fermented beef were closely related to the level of functonal proten damage. Reducton n proten occurred n a proten bands wth a molecular weght of 81.80 kD, 61.02 kD, 41.29 kD, 26.60 kD and 22.98 kD.

Table 1. Crude proten and proteolytc actvty of fermented beef

Parameters Slced fermented beef Ground fermented beef

Crude proten (% db) 81.77 ± 2.46a _{72.13 ± 2.84}b

Proteolytc actvty (U/g) 0.11 ± 0.04 0.09 ± 0.03

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These were due to faulty conformaton of the proten after fermentaton and proten structure changes nto a more smple peptdes and amno acds. Fadda et al. (1999) showed the degradaton of the proten bands because of proteolytc actvty of L. plantarum that occured n proten wth molecular weght 200 kD (myosn), 66 kD and 43 kD (actn). In ths research, molecular weght proten bands were (35 to 50 kD) hydrolyzed, whereas myosn and actn partally hydrolyzed after the noculaton of L. plantarum. These results further renforce the fact the degradaton of proten beef nto smpler form after fermentaton.

Amino acid composition of fermented beef

Amno acd composton of slced fermented beef was dfferent than ground fermented beef by descrptve analyss. Nonessental amno acds were detected by HPLC analyss of the aspartc acd, glutamc acd, serne, hstdn, glycne, argnne and alann, whle the essental amno acds detected were treonn, tyrosn, methonn, valn, phenlalann, soleusn, leusn and lysn. The quanttatve results can be seen n Table 2.

The results of HPLC analyss showed an ncrease n amno acd content after fermentaton. Ths occurred as a result of the proteolytc enzymes of L.plantarum

Table 2. Amno acd composton

Amno acd

Percentage of amno acd content (% w/w)

Increasng of amno acd n fermented beef compared than fresh beef (%)

Fresh DFD

Aspartc acd 1.66 2.93 2.56 76.50 54.22

Glutamc acd 3.42 5.64 4.79 64.91 40.06

Sern 0.79 1.31 1.09 65.82 37.97

Hstdn 0.70 1.02 0.91 45.71 30.00

Glysn 0.97 1.47 1.41 51.55 45,36

Threonn 0.98 1.58 1.27 61.22 29.59

Argnn 1.57 2.18 1.98 38.85 26.11

Alann 1.08 1.79 1.60 65.74 48.15

Tyrosn 0.67 0.99 0.79 47.46 17.91

Methonn 0.47 0.72 0.36 53.19 -23.40

Valn 0.90 1.46 1.48 62.22 64.44

Phenlalann 0.73 1.16 1.07 58.90 46.58

I – leusn 0.90 1.50 1.29 66.67 43.33

Leusn 1.48 2.40 2.09 62.16 41.22

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1B1. Fermentaton by L plantarum 1B1 could ncrease amno acd percentage of both fermented beef f compared to fresh DFD beef. Lactc acd produced from the fermentaton process caused a decrease n muscle pH beef. Ths declne contnued untl the pH reached ts optmum value. It then actvated proteolytc enzymes of

L.plantarum 1B1. The enzyme degraded Z-lne on myoflamen, elmnatng cross-brdge of actomyosn, and amno acd n actomyosn could be released. In general, amno acd percentages of slced fermented beef was hgher than ground fermented beef. It was supported by crude proten of slced fermented beef, that was hgher than ground fermented beef (Table 1).

Conclusons

Fermentaton by L. plantarum 1B1 on beef could ncrease crude proten of beef, whle slced fermented beef had a hgher proten content than ground fermented beef.

L. plantarum 1B1 had proteolytc actvty due to fermentaton. Fermentaton could degrade proten structure, as shown by reducton of proten bands wth a molecular weght of 81.80 kD, 61.02 kD, 41.29 kD, 26.60 kD and 22.98 kD. Fermentaton by L plantarum 1B1 could ncrease amno acd percentage of both fermented beef. Amno acd percentages of slced fermented beef was hgher than ground fermented beef. In general, slced fermented beef had better proten qualty than ground fermented beef.

Acknowledgement

Ths research was funded by Drectorate General of Hgher Educaton (DIKTI), Compettve Research Grant XII 2006.

References

AOAC. 2005. Offcal Methods of Analyss of AOAC Internatonal. 18th_{ed. Assoc.}

Off. Anal. Chem., Arlngton.

Aref, II. 2011. Characterzaton of Indgenous Lactc Acd Bactera from Beef as Probotc and Identfcaton by 16S rRNA gene sequencng. Dssertaton. Bogor Agrcultural Unversty.

Bergmeyer, H. U. and Karlfeld Gawehn. 1983. Methods of Enzymatc Analyss. Second Englsh Edton. Verlag Cheme Internatonal. Deerfeld Beach, Florda.

Fadda, S., G. Vgnolo, A. P. R Holgado and G. Olver. 1998. Proteolytc actvty1998. Proteolytc actvty of Lactobacillus strans solated from dry fermented sausages on muscle sarcoplasmc protens. J. Meat Scence (49) 1:11-18.

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Characterzaton of Muscle Sarcoplasmc and Myofbrllar Proten Hydrolyss Caused by Lactobacillus plantarum. Appled and Envronmental Mcrobology. 65: 3540-3546.