(....).hwp

연구기관 한국식품연구원 농림부

연구기관 한국식품연구원 농림부

Analytical Semi-preparative

Solution A 1) Solution B 2) Mixing Spreading Coating medium Corona treated OPP 3) film Casting Drying Room temp., 24 h

Sac I 5 Mature peda Deduced Factor Xa recognition sequence 3 Hind III Xba I Pst I Sal I EcoR I BamHI Xho I Kpn I Nde I Hind III Sac I

0.5Kb Ped + Sal I plasmid EcoRV Nde I peda pedb pedc pedd pucfp23 BamHI ped-hup-3 BamHI ped-hrp-3 pucpm5 BamHI ped-pmb-up Sac I ped-hrp pcwpm30 BamHI ped-pmb-up BamHI ped-hrp-3 pcwppm P32 ped-pmb-up ped-hrp-3 Primer

검체채취량

검체채취량

M 1 23.1 9.4 6.6 4.4 2.3 2.0 0.5

Kbp M 1 2 23.1 9.4 6.6 4.4 2.3 2.0 0.5

kbp M 1 2 3 4 23 9.4 6.6 4.4 2.3 2.0 0.5 1.2kb

kbp M 1 2 3 23 9.4 6.6 4.4 2.3 2.0 0.5 kbp M 1 2 23 9.4 6.6 4.4 2.3 2.0 0.5

kbp M 1 2 3 23 9.4 6.6 4.4 2.3 2.0 0.5 kbp M 1 2 23 9.4 6.6 4.4 2.3 2.0 0.5 1.1kbp

1 2

3 8 1200 7 1000 OD (at 660nm) 2 1 OD ph AU 6 5 ph 800 600 400 Activity(AU/mL) 4 200 0 0 3 6 9 12 15 18 21 24 Culture time(hr) 3 0 3 7 500 400 OD (at 660nm) 2 1 OD ph AU 6 5 ph 300 200 Activity(AU/mL) 100 0 0 3 6 9 12 15 18 21 24 27 30 33 36 39 Culture time(hr) 4 0

3 8 1200 OD (at 660nm) 2 1 OD ph AU 7 6 5 ph 1000 800 600 400 Pediocin activity(au/ml) 200 0 0 3 6 9 12 15 18 21 24 Culture time(hr) 4 0 3 8 1200 OD (at 660nm) 2 1 OD ph AU 7 6 5 ph 1000 800 600 400 Pediocin activity(au/ml) 200 0 0 3 6 9 12 15 Culture time(hr) 4 0

3 8 1200 7 1000 OD (at 660nm) 2 1 OD ph AU 6 5 ph 800 600 400 Activity(AU/mL) 4 200 0 0 3 6 9 12 15 18 21 24 Culture time(hr) 3 0 3.0 O.D(at 660nm) 2.5 2.0 1.5 1.0 ph 7.0 ph 6.8 ph 6.6 ph 6.4 ph 6.0 ph 5.8 ph 5.6 ph 5.4 0.5 0.0 0 3 6 9 12 15 Culture time(hr)

7.5 ph 7.0 6.5 6.0 5.5 ph 7.0 ph 6.8 ph 6.6 ph 6.4 ph 6.0 ph 5.8 ph 5.6 ph 5.4 5.0 4.5 4.0 0 3 6 9 12 15 Culture time(hr) 1600 Pediocin activity (AU/mL) 1400 1200 1000 800 600 400 ph 7.0 ph 6.8 ph 6.6 ph 6.4 ph 6.0 ph 5.8 ph 5.6 ph 5.4 200 0 0 3 6 9 12 15 Culture time(hr)

3.0 O.D (at 660nm) 2.5 2.0 1.5 1.0 Peptone Tryptone Meat extract Yeast extract 0.5 0.0 0 3 6 9 12 15 18 Culture time(hr) 6.5 6.0 Peptone Tryptone Meat extract Yeast extract 5.5 ph 5.0 4.5 4.0 0 3 6 9 12 15 18 Culture time(hr)

1200 Pediocin activity(au/ml) 1000 800 600 400 Peptone Tryptone Meat extract Yeast extract 200 0 0 3 6 9 12 15 18 Culture time(hr) 1400 Pediocin activity(au/ml) 1200 1000 800 600 400 0.01% 0.02% 0.04% 0.06% 0.10% 200 0 9 12 15 Culture time(hr)

1400 Pediocin activity(au/ml) 1200 1000 800 600 400 0.005% 0.015% 0.025% 0.035% 0.045% 200 0 9 12 15 Culture time(hr)

2.5 2.0 O.D. at 660nm 1.5 1.0 0.5 control whey 10% whey 2% whey 0.2% 0.0 0 12 24 36 48 Incubation time(hr) 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 control tryp1%+whey10% tryp1%+whey2% tryp1%+whey0.2% tryp2%+whey10% tryp2%+whey2% tryp2%+whey0.2% tryp3%+whey10% tryp3%+whey2% tryp3%+whey0.2% 0.0 0 12 24 36 48 Incubation time(hr)

2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 control yeast1%+whey10% yeast1%+whey2% yeast1%+whey0.2% yeast2%+whey10% yeast2%+whey2% yeast2%+whey0.2% yeast3%+whey10% yeast3%+whey2% yeast3%+whey0.2% 0.0 0 12 24 36 48 Incubation time(hr) 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 control beef1%+whey10% beef1%+whey2% beef1%+whey0.2% beef2%+whey10% beef2%+whey2% beef2%+whey0.2% beef3%+whey10% beef3%+whey2% beef3%+whey0.2% 0.0 0 12 24 36 48 Incubation time(hr)

3.0 2.5 O.D. at 660nm 2.0 1.5 1.0 control whey2%+pep1% whey2%+pep2% whey2%+pep3% whey2%+pep4% whey2%+pep5% whey2%+pep6% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) 3.0 O.D. at 660nm 2.5 2.0 1.5 1.0 control whey4%+pep1% whey4%+pep2% whey4%+pep3% whey4%+pep4% whey4%+pep5% whey4%+pep6% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr)

3.0 O.D. at 660nm 2.5 2.0 1.5 1.0 control whey6%+pep1% whey6%+pep2% whey6%+pep3% whey6%+pep4% whey6%+pep5% whey6%+pep6% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) O.D. at 660nm 3.0 2.5 2.0 1.5 1.0 control whey2%+pep6% whey3%+pep6% whey4%+pep6% whey6%+pep6% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr)

3.0 O.D. at 660nm 2.5 2.0 1.5 1.0 control whey4%+pep1% whey4%+pep2% whey4%+pep3% whey4%+pep4% whey4%+pep5% whey4%+pep6% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr)

O.D. at 660nm 3.0 2.5 2.0 1.5 1.0 control yeast1%+whey2% yeast2%+whey2% yeast3%+whey2% yeast4%+whey2% yeast5%+whey2% yeast6%+whey2% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) O.D. at 660nm 3.0 2.5 2.0 1.5 1.0 control yeast1%+whey4% yeast2%+whey4% yeast3%+whey4% yeast4%+whey4% yeast5%+whey4% yeast6%+whey4% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr)

O.D. at 660nm 3.0 2.5 2.0 1.5 1.0 control yeast1%+whey6% yeast2%+whey6% yeast3%+whey6% yeast4%+whey6% yeast5%+whey6% yeast6%+whey6% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) 3.0 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 control yeast6%+whey2% yeast6%+whey4% yeast6%+whey6% 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr)

3.0 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 control yeast0%+glc0% yeast0%+glc1% yeast0%+glc2% yeast0%+glc3% 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) O.D. at 660nm 3.0 2.5 2.0 1.5 1.0 control yeast1%+glc0% yeast1%+glc1% yeast1%+glc2% yeast1%+glc3% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr)

O.D. at 660nm 3.0 2.5 2.0 1.5 1.0 control yeast2%+glc0% yeast2%+glc1% yeast2%+glc2% yeast2%+glc3% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) O.D. at 660nm 3.0 2.5 2.0 1.5 1.0 control yeast3%+glc0% yeast3%+glc1% yeast3%+glc2% yeast3%+glc3% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr)

O.D. at 660nm 3.0 2.5 2.0 1.5 1.0 control yeast4%+glc0% yeast4%+glc1% yeast4%+glc2% yeast4%+glc3% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) O.D. at 660nm 3.0 2.5 2.0 1.5 1.0 control yeast5%+glc0% yeast5%+glc1% yeast5%+glc2% yeast5%+glc3% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr)

O.D. at 660nm 3.0 2.5 2.0 1.5 1.0 control yeast0%+glc1% yeast1%+glc1% yeast2%+glc1% yeast3%+glc1% yeast4%+glc1% yeast5%+glc1% yeast6%+glc1% 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) 3.0 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) control yeast6%+glc1% yeast3%+pep3%+glc1% glc1%+pep1% glc1%+pep2% glc1%+pep3% glc1%+pep4% glc1%+pep5% glc1%+pep6%

3.0 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) control yeast6%+glc1% yeast3%+pep3%+glc1% glc2%+pep1% glc2%+pep2% glc2%+pep3% glc2%+pep4% glc2%+pep5% glc2%+pep6% 3.0 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) control yeast6%+glc1% yeast3%+pep3%+glc1% glc3%+pep1% glc3%+pep2% glc3%+pep3% glc3%+pep4% glc3%+pep5% glc3%+pep6%

3.0 2.5 O.D. at 660bn 2.0 1.5 1.0 0.5 control yeast6%+glc1% yeast3%+pep3%+glc1% glc2%+pep1% 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) 3.0 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 control yeast0% yeast6% yeast3%+tryp1% yeast3%+tryp2% yeast3%+tryp3% yeast3%+beef1% yeast3%+beef2% yeast3%+beef3% 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr)

3.0 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 control yeast0% yeast6% yeast3%+tryp1% yeast3%+tryp2% yeast3%+tryp3% 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) 3.0 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 control yeast0% yeast6% yeast3%+beef1% yeast3%+beef2% yeast3%+beef3% 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr)

3.0 2.5 O.D. at 660nm 2.0 1.5 1.0 0.5 0.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) 0.1% 0.2% 0.4% 0.6% 0.8% 1.0% 1.5% 2.0% 3.0% 4.0% 5.0% 3.0 7.0 2000 O.D. at 660nm 2.5 2.0 1.5 1.0 0.5 6.5 6.0 5.5 5.0 4.5 ph 1500 1000 500 Activity(AU/mL) 0.0 4.0 0 3 6 9 12 15 18 21 24 Incubation time(hr) 0

OD (at 660nm 1.0 0.8 0.6 0.4 0.2 Untreated Pediocin 50ppm Pediocin 100ppm Pediocin 250ppm Pediocin 25ppm Pediocin 75ppm Pediocin 125ppm 0.0 0 2 4 6 8 10 Incubation time(hr) 1.0 OD (at 660nm 0.8 0.6 0.4 Untreated Polylysine 50ppm Polylysine 100ppm Polylysine 250ppm Polylysine 25ppm Polylysine 75ppm Polylysine 125ppm 0.2 0.0 0 2 4 6 8 10 Incubation time(hr)

OD (at 660nm) 1.0 0.8 0.6 0.4 Untreated Nis in 50ppm Nis in 100ppm Nis in 250ppm Nisin 25ppm Nisin 75ppm Nis in 125ppm 0.2 0.0 0 2 4 6 8 10 Incubation time(hr) OD (at 660nm 1.4 1.2 1.0 0.8 0.6 0.4 0.2 Untreated Pediocin 50ppm Pediocin 100ppm Pediocin 250ppm Pediocin 25ppm Pediocin 75ppm Pediocin 125ppm 0.0 0 2 4 6 8 10 12 Incubation time(hr)

1.0 0.8 Untreated Polylysine 50ppm Polylysine 100ppm Polylysine 250ppm Polylysine 25ppm Polylysine 75ppm Polylysine 125ppm OD (at 660nm) 0.6 0.4 0.2 0.0 0 2 4 6 8 10 12 Incubation time(hr) 1.0 OD (at 660nm) 0.8 0.6 0.4 Untreated Nisin 50ppm Nisin 100ppm Nisin 250ppm Nisin 25ppm Nisin 75ppm Nisin 125ppm 0.2 0.0 0 2 4 6 8 10 12 Incubation time(hr)

OD (at 660nm) 1.2 1.0 0.8 0.6 0.4 Untreated Pediocin 50ppm Pediocin 100ppm Pediocin 250ppm Pediocin 25ppm Pediocin 75ppm Pediocin 125ppm 0.2 0.0 0 2 4 6 8 10 12 Incubation time(hr) OD (at 660nm) 1.0 0.8 0.6 0.4 0.2 Untreated Polylysine 50ppm Polylysine 100ppm Polylysine 250ppm Polylysine 25ppm Polylysine 75ppm Polylysine 125ppm 0.0 0 2 4 6 8 10 12 Incubation time(hr)

OD (at 660nm) 1.0 0.8 0.6 0.4 Untreated Nisin 50ppm Nisin 100ppm Nisin 250ppm Nisin 25ppm Nisin 75ppm Nisin 125ppm 0.2 0.0 0 2 4 6 8 10 12 Incubation time(hr) OD (at 660nm) 1.0 0.8 0.6 0.4 Untreated Pediocin 25ppm Pediocin 50ppm Pediocin 75ppm Pediocin 100ppm Pediocin 125ppm Pediocin 250ppm 0.2 0.0 0 2 4 6 8 10 Incubation time(hr)

OD (at 660nm) 1.0 0.8 0.6 0.4 Untreated Polylysine 25ppm Polylysine 50ppm Polylysine 75ppm Polylysine 100ppm Polylysine 125ppm Polylysine 250ppm 0.2 0.0 0 2 4 6 8 10 Incubation time(hr) OD (at 660nm) 1.4 1.2 1.0 0.8 0.6 0.4 0.2 Untreated Nisin 50ppm Nisin 100ppm Nisin 250ppm Nisin 25ppm Nisin 75ppm Nisin 125ppm 0.0 0 2 4 6 8 10 Incubation time(hr)

OD (at 660nm) 1.0 0.8 0.6 0.4 Untreated Pediocin 25ppm Pediocin 50ppm Pediocin 75ppm Pediocin 100ppm Pediocin 125ppm Pediocin 250ppm 0.2 0.0 0 2 4 6 8 10 Incubation time(hr) OD (at 660nm) 1.0 0.8 0.6 0.4 Untreated Polylysine 25ppm Polylysine 50ppm Polylysine 75ppm Polylysine 100ppm Polylysine 125ppm Polylysine 250ppm 0.2 0.0 0 2 4 6 8 10 Incubation time(hr)

OD (at 660nm) 1.0 0.8 0.6 0.4 0.2 Untreated Nisin 50ppm Nisin 100ppm Nisin 250ppm Nisin 25ppm Nisin 75ppm Nisin 125ppm 0.0 0 2 4 6 8 10 Incubation time(hr)

log cfu/g 12 10 8 6 4 Untreated Pediocin 25ppm Pediocin 100ppm Pediocin 250ppm Pediocin 500ppm Pediocin 1000ppm 2 0 0 3 6 Storage time(days) log cfu/g 10 8 6 4 Untreated Polylysine 25ppm Polylysine 250ppm Polylysine 1000ppm Polylysine 4000ppm Polylysine 8000ppm 2 0 0 3 6 Storage time(days)

10 8 Untreated Nis in 100ppm Nis in 500ppm Nisin 25ppm Nis in 250ppm Nis in 1000ppm log cfu/g 6 4 2 0 0 3 6 Storage time(days) 80.00 L value 75.00 Un tre ate d P ediocin 100ppm P ediocin 500ppm Pediocin 25ppm Pediocin 250ppm P ediocin 1000ppm 70.00 0 3 6 Storage time(days)

85.00 80.00 L value 75.00 70.00 Un tre a te d Polylysine 250ppm Polylysine 4000ppm Polylysine 25ppm P olylys in e 1000ppm P olylys in e 8000ppm 65.00 0 3 6 Storage time(days) 85.00 80.00 L value 75.00 70.00 Untreated Nis in 100ppm Nis in 500ppm Nisin 25ppm Nis in 250ppm Nis in 1000ppm 65.00 0 3 6 Storage time(days)

10 8 Untreated Polylysine 0.2% Polylysine 0.4% Polylysine 0.6% Polylysine 0.8% Polylysine 1.0% Polylysine 1.5% Log cfu/g 6 4 2 0 0 3 6 Storage time(days)

85 80 L value 75 70 65 60 Untreated Polylysine 0.2% Polylysine 0.4% Polylysine 0.6% Polylysine 0.8% Polylysine 1.0% Polylysine 1.5% 0 3 6 Storage time(days)

ogcfu/10 8 6 Untreated 100 ppm nisin 4 200 ppm nisin 400 ppm nisin 2 800 ppm nisin 1200 ppm nisinl g log CFU/g 0 0 3 6 Storage time(days) 80 75 L value 70 65 60 Untreated 100 ppm nisin 200 ppm nisin 400 ppm nisin 800 ppm nisin 1200 ppm nisin 0 3 6 Storage time(days)

ogcfu/10 8 6 Untreated 4 100 ppm pediocin 200 ppm pediocin 1200 ppm 400 ppm pediocin 2 pediocinl800 ppm pediocin 0 0 3 6 Storage time(days) g log CFU/g 80 75 L value 70 65 60 Untreated 100 ppm pediocin 200 ppm pediocin 400 ppm pediocin 800 ppm pediocin 1200 ppm pediocin 0 3 6 Storage time(days)

10 8 Log cfu/g 6 4 2 Untreated Lysozyme 0.01% Lysozyme 0.05% Lysozyme 0.1% Lysozyme 0.3% Lysozyme 0.5% 0 0 3 6 Storage time(days) L value 85 80 75 70 Untreated Lysozyme 0.01% Lysozyme 0.05% Lysozyme 0.1% Lysozyme 0.3% Lysozyme 0.5% 65 60 0 3 6 Storage time(days)

85 80 75 Untreated Chito-oligo 0.05% Chito-oligo 0.1% Chito-oligo 0.3% L value 70 65 60 55 0 3 6 Storage time(days) 10 8 Log cfu/g 6 4 2 0 Untreated Chito-oligo 0.05% Chito-oligo 0.1% Chito-oligo 0.3% 0 3 6 Storage time(days)

10 8 Log cfu/g 6 4 2 Untreated GSE 250ppm GSE 1000ppm GSE 50ppm GSE 500ppm 0 0 3 6 Storage time(days) L value 90 85 80 75 70 65 60 Untreated GSE 250ppm GSE 1000ppm GSE 50ppm GSE 500ppm 55 50 0 3 6 Storage time(days)

ogcfu/10 8 6 4 Untreated 100 ppm vitaten-s 2 1000 ppm vitaten-s 10000 ppm vitaten-sl g log CFU/g 0 0 3 6 Storage time(days) 75 70 L value 65 60 Untreated 100 ppm vitaten-s 1000 ppm vitaten-s 10000 ppm vitaten-s 55 0 3 6 Storage time(days)

ogcfu/10 8 6 Untreated 4 50 ppm hinokitiol 100 ppm hinokitiol 2 150 ppm hinokitiol 200 ppm hinokitioll g log CFU/g 0 0 3 6 Storage time(days) 85 L value 80 75 70 Untreated 50 ppm hinokitiol 100 ppm hinokitiol 150 ppm hinokitiol 200 ppm hinokitiol 0 3 6 Storage time(days)

olog CFU/glgCFU/10 8 6 4 2 0 g 0 3 6 Storage time(days)

90 85 L value 80 75 70 65 0 3 6 Storage time(days)

10 8 log cfu/g Log 6 4 2 0 Untreated D W Citric acid 0.1% Citric acid 0.25% Citric acid 0.5% 8 log cfu/g Log 6 4 2 0 Untreated Citric acid 0.75% Citric acid 1.0% Citric acid 1.25% Citric acid 1.5% 0 3 6 Storage time(days)

80 75 70 L value 65 60 55 Untreated D W Citric acid 0.1% Citric acid 0.25% Citric acid 0.5% 75 70 L value 65 60 55 50 Untreated Citric acid 0.75% Citric acid 1.0% Citric acid 1.25% Citric acid 1.5% 0 3 6 Storage time(days)

8 log cfu/g Log 6 4 2 0 8 Untreated Malic acid 0.1% Malic acid 0.25% Malic acid 0.5% log cfu/g Log 6 4 2 0 Untreated Malic acid 0.75% Malic acid 1.0% Malic acid 1.25% Malic acid 1.5% 0 3 6 Storage time(days)

85 80 75 L value 70 65 60 55 Untreated Malic acid 0.1% Malic acid 0.25% Malic acid 0.5% 75 70 L value 65 60 55 50 45 Untreated Malic acid 0.75% Malic acid 1.0% Malic acid 1.25% Malic acid 1.5% 0 3 6 Storage time(days)

10 8 Treat. I Treat. III Treat. V Treat. VII Treat. II Treat. IV Treat. VI Log cfu/g 6 4 2 0 8 Treat. VIII Treat. X Treat. XII Treat. IX Treat. XI Treat. XIII Log cfu/g 6 4 2 0 0 3 6 Storage time(days)

85 80 Treat. I Treat. III Treat. V Treat. VII Treat. II Treat. IV Treat. VI L value 75 70 65 80 Treat. VIII Treat. X Treat. XII Treat. IX Treat. XI Treat. XIII L value 75 70 65 0 3 6 Storage time(days)

10 8 log cfu/g Log 6 4 2 0 D W normal temp., 1min D W 40, 1 min D W 40, 2 min D W 50, 30 sec D W 50, 1 min D W 50, 2 min 8 log cfu/g Log 6 4 2 0 D W normal temp., 1min D W 60, 30 sec D W 60, 1 min D W 60, 2 sec D W 70, 1 min 0 3 6 Storage time(days)

L value 85 80 75 70 65 60 55 50 D W normal temp., 1min D W 40, 2 min D W 50, 1 min D W 40, 1 min D W 50, 30 sec D W 50, 2 min L value 75 70 65 60 55 50 45 40 D W normal temp., 1min D W 60, 1 min D W 60, 30 sec D W 70, 1 min D W 60, 2 sec 0 3 6 Storage time(days)

10 8 Log cfu/g 6 4 2 0 0 3 6 Storage time(days) Untreated Ascorbic acid 0.1% Ascorbic acid 0.25% Ascorbic acid 0.5% Ascorbic acid 0.75% Ascorbic acid 1.0%

L value 80 75 70 Untreated Ascorbic acid 0.1% Ascorbic acid 0.25% Ascorbic acid 0.5% Ascorbic acid 0.75% Ascorbic acid 1.0% 65 60 0 3 6 Storage time(days) 10 8 Log cfu/g 6 4 2 0 0 3 6 Storage time(days) Untreated Calcium chloride 0.1% Calcium chloride 0.25% Calcium chloride 0.5% Calcium chloride 0.75% Calcium chloride 1.0%

80 75 L value 70 65 60 55 Untreated Calcium chloride 0.1% Calcium chloride 0.25% Calcium chloride 0.5% Calcium chloride 0.75% Calcium chloride 1.0% 0 3 6 Storage time(days)

10 8 Untreated Treat. Ⅱ Treat. Ⅳ Treat. Ⅰ Treat. Ⅲ Log cfu/g 6 4 2 0 0 5 10 15 20 Storage time(days)

90 85 80 L value 75 70 65 60 Untreated Treat. Ⅰ Treat. Ⅱ Treat. Ⅲ Treat. Ⅳ 0 5 10 15 20 Storage time(days)

5 4 Hardness(kg 3 2 1 Untreated Treat. Ⅱ Treat. Ⅳ Treat. Ⅰ Treat. Ⅲ 0 0 5 10 15 20 S torage time(days)

0.50 0.40 Un tre ate d Treat. Ⅱ Treat. Ⅳ Treat. Ⅰ Treat. Ⅲ Weight loss(% 0.30 0.20 0.10 0.00 0 5 10 15 20 S torage time(days)

Titratable acidity(% 0.7 0.6 0.5 0.4 0.3 0.2 0.1 Untreated Treat. Ⅱ Treat. Ⅳ Treat. Ⅰ Treat. Ⅲ 0.0 0 10 20 Storage time(days)

ogcfu/gogcfu/10 8 Untreated -- -- H treat. log CFU/ NP treat. -- -- HNP treat.l6 4 2 0 0 5 10 15 20 Storage time(days) g l

85 80 Untreated NP treat. -- -- H treat. -- -- HNP treat. L value 75 70 65 60 0 5 10 15 20 Storage time(days)

5 4 Force(kg 3 2 1 Untreated NP treat. -- -- H treat. -- -- HNP treat. 0 0 10 20 Storage time(days)

0.4 Untreated H treat. Titratable acidity( 0.3 0.2 0.1 NP treat. HNP treat. 0.0 0 10 20 Storage time(days)

56 37 Weissella koreensis AY035892 Weissella viridescens AB023236 83 87 Weissella confusa AB023241 Weissella cibaria AF312874 Weissella soli AY028260 79 49 Lactobacillus reuteri X76328 100 Lactobacillus vaginalis AF243177 56 47 Lactobacillus colehominis AJ292530 Weissella thailandensis AB0238 Weissella hellenica AB015642 Weissella paramesenteroides S67831 B193 0.05

56 Weissella koreensis AY035892 Weissella viridescens AB023236 83 87 Weissella confusa AB023241 Weissella cibaria AF312874 Weissella soli AY028260 56 79 100 53 Lactobacillus colehominis AJ292530 Lactobacillus vaginalis AF243177 Lactobacillus reuteri X76328 Weissella thailandensis AB0238 Weissella hellenica AB015642 Weissella paramesenteroides S67831 B193

1.2 OD (at 660nm) 1.0 0.8 0.6 0.4 E. coli KFRI 242 E. coli KFRI 242 + Ped M E. coli KFRI 242 + 2774 E. coli KFRI 242 + 2775 E. coli KFRI 242 + 2777 0.2 0.0 0 1 2 3 4 5 6 7 8 Cultivation time (hr) 1.0E +10 1.0E +09 Total count[log(cfu/ml)] 1.0E +08 1.0E +07 1.0E +06 1.0E +05 U n tre a te d P. acidilactici 2774 2775 2777 1.0E +04 0 1 2 3 5 7 Storage time(day)

1.0E +10 Lactic acid bacteria count[log(cfu/ml)] 1.0E +09 1.0E +08 1.0E +07 1.0E +06 1.0E +05 1.0E +04 U n tre a te d 2774 2775 2777 P. acidilactici 0 1 2 3 5 7 Storage time(day) 1.0E +05 Coliform bacterial count[log(cfu/ml)] 1.0E +04 1.0E +03 1.0E +02 1.0E +01 1.0E +00 U n tre a te d 2774 2775 2777 P. acidilactici 0 1 2 3 5 7 Storage time(day)

7 6 5 ph 4 3 Untreated P. acidilactici 2 1 2774 2775 2777 0 0 1 2 3 5 7 Storage time(day) 1.0 U n tre a te d P. acidilactici 0.8 2774 2775 2777 Acidity(%) 0.6 0.4 0.2 0.0 0 1 2 3 5 7 Storage time(day)

Microbial count[log(cfu/ml) 1.0E+09 1.0E+08 1.0E+07 Untreated Polylysine 250ppm Mixture Ped M 250ppm Nisin 250ppm 1.0E+06 0 1 2 3 6 9 12 Storage time(day)

1.0E+09 Untreated Ped M 250ppm Polylysine 250ppm Nis in 250ppm Coliform count[log(cfu/ml)] 1.0E+08 1.0E+07 Mixture 1.0E+06 0 1 2 3 6 9 12 Storage time(day) 1.0E+09 Lactic acid bacterial count[log(cfu/ml)] 1.0E+08 1.0E+07 Untreated Polylysine 250ppm Mixture Ped M 250ppm Nisin 250ppm 1.0E+06 0 1 2 3 6 9 12 Storage time(day)

1.0E+09 Yeast & Mold count[log(cfu/ml)] 1.0E+08 1.0E+07 Untreated Polylysine 250ppm Mixture Ped M 250ppm Nisin 250ppm 1.0E+06 0 1 2 3 6 9 12 Storage ti me(day) 4.00 3.90 3.80 ph 3.70 3.60 Untreated Polylysine 250ppm Mixture Ped M 250ppm Nisin 250ppm 3.50 0 1 2 3 6 9 12 Storage time(day)

1.2 1.0 0.8 Acidity(%) 0.6 0.4 0.2 Untreated P oly ly s ine 250ppm Mixture P e d M 250ppm Nis in 250ppm 0.0 0 1 2 3 6 9 12 Storage time(day) 60 50 Amino nitrogen(mg%) 40 30 20 10 Untreated Polylysine 250ppm Mixture Ped M 250ppm Nisin 250ppm 0 0 1 2 3 6 9 12 Storage time(day)

Bacterial species Antimicrobial materials Gram Pediocin Nisin staining A 1) B 2) A 1) B 2) Bacillus cereus KCTC 1012 + - - - - " KCTC 1092 + - - - - " KCTC 1094 + - - - - " KCTC 1526 + - - - + " KCTC 1093 + + + - - " KCTC 3711 + - - - - " KCTC 3674 + + + - - " KCCM 40133 + - - - - " KCCM 40138 + - - - - " KCCM 11341 + +++ +++ - - Lactobacillus plantarum KFRI 00466 + - - + + " KFRI 00402 + - - - - " KFRI 00402 + - - - - " KFRI 00402 + - - - - Lactobacillus acidophilus KFRI 00340 + - - - - " KFRI 00162 + + ++ + ++ Bacillus subtilis KFRI 01120 + + + - - " KFRI 01121 + - - - - " KFRI 01125 + - - + ++ " KFRI 00119 + - - + ++ Lactobacillus brevis KFRI 00805 + - - + ++ " KFRI 00806 + + + + +++ " KFRI 00353 + - - + ++ " KFRI 00221 + - - + ++ Enterococcus faecalis KFRI 00354 + + + + + Klebsiella pneumoniae KCTC 1726 - - - - - " KCTC 2241 - - - - - " KCTC 2242 - - - - - " KCTC 2243 - - - - - " KCTC 2244 - + + - - " KCTC 2245 - + + - - " KCTC 2246 - - - - - " KCTC 2248 - - - - -

4.8 4.7 Control 0.03% 0.05% 0.10% ph 4.6 4.5 4.4 0 24 48 72 96 Storage time (hr) 7 Total count(log. CFU/mL) 6 5 4 3 2 Control 0.03% 1 0.05% 0 0.10% 0 24 48 72 96 Storage time (hr)

Coliform count(cfu/ml) 350 300 250 200 150 100 50 0 Control 0.03% 0.05% 0.10% 0 24 48 72 96 Storage time (hr)

Yeast&Mold count(log. CFU/mL) 6 5 4 3 2 Control 0.03% 1 0.05% 0.10% 0 0 24 48 72 96 Storage time (hr)

8 6 ph 4 Control 2 0.03% 0.05% 0.10% 0 0 36 72 108 144 180 Storage time (hr) 30 25 20 Control 0.03% 0.05% 0.10% Acidity 15 10 5 0 0 36 72 108 144 180 Storage time (hr)

Turbidity(NTU) 1000 800 600 400 200 Control 0.03% 0.05% 0.10% 0 0 36 72 108 144 180 Storage time (hr)

Total count(log. CFU/mL) 10 Control 8 0.03% 0.05% 6 0.10% 4 2 0 0 36 72 108 144 180 Storage time (hr)

Coliform count(cfu/ml) 3500 3000 2500 2000 1500 1000 500 0 Control 0.03% 0.05% 0.10% 0 36 84 132 168 204 Storage time (hr)

6.9 6.8 6.7 ph 6.6 6.5 Control 0.03% 6.4 0.05% 0.10% 6.3 0 24 48 72 96 120 144 168 Storage time (hr)

7 Total count(log. CFU/mL) 6 5 Control 4 0.03% 0.05% 0.10% 3 0 24 48 72 96 120 144 168 Storage time (hr) Coilform count(log. CFU/mL) 8 6 4 Control 2 0.03% 0.05% 0.10% 0 0 24 48 72 96 120 144 168 Storage time (hr)

ogcfu/10 Untreated -- -- H treat. 8 NP treat. -- -- HNP treat.llog CFU/ 6 4 2 0 0 1 2 3 4 Storage time(days) g

ogcfu/6 5 Untreated -- -- H treat. NP treat. -- -- HNP 4 3 2 treat.l g log CFU/ 1 0 0 1 2 3 4 Storage time(days)

150 125 Untreated H treat. Force(g) 100 75 50 NP treat. HNP treat. 25 0 0 2 4 Storage time(days)

0.8 OD at 660nm 0.6 0.4 0.2 0.0 0 3 6 9 15 18 21 24 Incubation time(h) 0.8 OD at 660nm 0.6 0.4 0.2 0.0 0 3 6 9 15 18 21 24 Incubation time(h)

0.8 OD at 660nm 0.6 0.4 0.2 0.0 0 3 6 9 15 18 21 24 Incubation time(h) 25 Polylysine concentration( μg /ml) 20 15 10 5 ` 0 0 1 2 3 4 5 6 7 Storage time(days)

8 Microbial count[log(cfu/ml)] 6 4 2 0 0 1 2 3 4 5 6 7 Incubation time(days) 8 Microbial count[log(cfu/ml)] 6 4 2 0 0 1 3 5 7 Incubation time(days)

25 Polylysine concentration( μg /ml) 20 15 10 5 0 1 3 5 Storage time(days)

8 Microbial count[log(cfu/ml)] 6 4 2 0 0 1 3 5 7 Incubation time(days) 8 Microbial count[log(cfu/ml)] 6 4 2 0 0 1 3 5 7 Incubation time(days)

25 Polylysine concentration( μg /ml) 20 15 10 5 0 1 3 5 Storage time(days) 8 Microbial count[log(cfu/ml)] 6 4 2 0 0 1 3 5 7 Incubation time(days)

Microbial count[log(cfu/ml)] 8 6 4 2 0 0 1 3 5 7 Incubation time(days)

Microbial count[log(cfu/ml)] 8 6 4 2 0 0 1 3 5 7 Incubation time(days)

Microbial count[log(cfu/ml)] 8 6 4 2 0 0 1 3 5 7 Incubation time(days)

10 Microbial count[log(cfu/g)] 8 6 4 2 0 0 2 4 6 8 10 Storage time(days)

10 Coliform count[log(cfu/g)] 8 6 4 2 0 0 2 4 6 8 10 Storage time(days)

6.0 4.5 Overall acceptability 3.0 1.5 0.0 0 2 4 6 8 10 Storage time(days)