inhibition activity on angiotensin converting … filecontains lactic acid bacteria and khamir....

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Seminar Nasional Sains dan Teknologi (Senastek) IV, Bali, Indonesia 2017 1 INHIBITION ACTIVITY ON ANGIOTENSIN CONVERTING ENZYME OF CONSUMPTIVE KEFIR DURING DIFFERENT LENGTH OF STORAGE Lindawati, S. A., N. G. K. Roni And I.D.K. Harya Putra Fakulty of Animal Science, Udayana University JL.PB. Sudirman Denpasar e-mail: [email protected] Abstract Consumptive kefir is milk product made by addition of kefir seed as starter. Kefir seed contains lactic acid bacteria and khamir. Lactic acid bacteria may stimuli the production of bioactive peptide in the consumptive kefir. Due to the present of this bioactive peptide, then the consumptive kefir is regarded as functional food stuff as it is beneficial for health status. One of its beneficial effects on health status can be related to its prevention to hypertension as it may inhibit activity of Angiotensin Converting Enzyme (ACE). Thus, the present research work reported in this paper was aimed to determine the chemical quality (protein, fat, lactose, pH and the total acid contents), microbiological quality and inhibition activity on ACE during different length of storage of the consumptive kefir. A Completely Randomized Design (CRD) was applied with 5 treatments and 3 replications. The treatments included different length of storage of the consumptive kefir for 0, 3, 6, 9 and 12 days at temperature of 4 o C. As a conclusion, consumptive kefir stored for 12 days has the highest inhibition activity on ACE, that was 39.32%, IC 50 was 0.98 mg/ml with protein content 4.09; fat: 2.01; lactose: 3.41; total acid: 2.95; and pH: 3.57, and the total lactic acid bacteria was 2,82 x 10 6 cfu/g 2,39 x 10 8 cfu/g and no growth of E. coli bacteria was noted. Keywords: consumptive kefir, angiotensin converting enzyme (ACE), length of storage

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Page 1: INHIBITION ACTIVITY ON ANGIOTENSIN CONVERTING … filecontains lactic acid bacteria and khamir. Lactic acid bacteria may stimuli the production of bioactive peptide in the consumptive

Seminar Nasional Sains dan Teknologi (Senastek) IV, Bali, Indonesia 2017

1

INHIBITION ACTIVITY ON ANGIOTENSIN CONVERTING

ENZYME OF CONSUMPTIVE KEFIR DURING DIFFERENT

LENGTH OF STORAGE

Lindawati, S. A., N. G. K. Roni And I.D.K. Harya Putra

Fakulty of Animal Science, Udayana University

JL.PB. Sudirman Denpasar

e-mail: [email protected]

Abstract

Consumptive kefir is milk product made by addition of kefir seed as starter. Kefir seed

contains lactic acid bacteria and khamir. Lactic acid bacteria may stimuli the production of

bioactive peptide in the consumptive kefir. Due to the present of this bioactive peptide, then the

consumptive kefir is regarded as functional food stuff as it is beneficial for health status. One of

its beneficial effects on health status can be related to its prevention to hypertension as it may

inhibit activity of Angiotensin Converting Enzyme (ACE). Thus, the present research work

reported in this paper was aimed to determine the chemical quality (protein, fat, lactose, pH and

the total acid contents), microbiological quality and inhibition activity on ACE during different

length of storage of the consumptive kefir. A Completely Randomized Design (CRD) was applied

with 5 treatments and 3 replications. The treatments included different length of storage of the

consumptive kefir for 0, 3, 6, 9 and 12 days at temperature of 4oC. As a conclusion, consumptive

kefir stored for 12 days has the highest inhibition activity on ACE, that was 39.32%, IC 50 was

0.98 mg/ml with protein content 4.09; fat: 2.01; lactose: 3.41; total acid: 2.95; and pH: 3.57, and

the total lactic acid bacteria was 2,82 x 106 cfu/g – 2,39 x 10 8 cfu/g and no growth of E. coli

bacteria was noted.

Keywords: consumptive kefir, angiotensin converting enzyme (ACE), length of storage

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PENELITIAN PRODUK TERAPANPENELITIAN PRODUK TERAPAN

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AKTIVITAS PENGHAMBATAN ANGIOTENSINCONVERTING ENZIM DARI KEFIR KONSUMSI

SELAMA PENYIMPANAN BERBEDA

AKTIVITAS PENGHAMBATAN ANGIOTENSINCONVERTING ENZIM DARI KEFIR KONSUMSI

SELAMA PENYIMPANAN BERBEDA

www.themegallery.com

Ir.

Ir. Sri Anggreni Lindawati, MSiIr. Ni Gusti Ketut Roni, M.Si

Prof. Dr. Ir. D.K. Harya Putra,M.Si

Ir.

Ir. Sri Anggreni Lindawati, MSiIr. Ni Gusti Ketut Roni, M.Si

Prof. Dr. Ir. D.K. Harya Putra,M.Si

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SUSU PRODUK HASIL TERNAK YANGBERNUTRISI TINGGI

FERMENTASIFUNGSIONAL

KEFIR KONSUMSI:- Susu yg

difermentasidengan biji kefir(BAL), yang

sebelumdikonsumsi,

disaringterlebih dahulu(tidak mengandungbiji kefir)

- KAUKASIUS- Di Bali sedikit

informasi

KEFIR KONSUMSI:- Susu yg

difermentasidengan biji kefir(BAL), yang

sebelumdikonsumsi,

disaringterlebih dahulu(tidak mengandungbiji kefir)

- KAUKASIUS- Di Bali sedikit

informasi

PENDAHULUANPENDAHULUAN

KEFIR KONSUMSI:- Susu yg

difermentasidengan biji kefir(BAL), yang

sebelumdikonsumsi,

disaringterlebih dahulu(tidak mengandungbiji kefir)

- KAUKASIUS- Di Bali sedikit

informasi

KEFIR KONSUMSI:- Susu yg

difermentasidengan biji kefir(BAL), yang

sebelumdikonsumsi,

disaringterlebih dahulu(tidak mengandungbiji kefir)

- KAUKASIUS- Di Bali sedikit

informasi

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Foto kefirKESEHATAN:antihipertensi

- BIJI KEFIR……BAL- FUNGSI BAL- PEPTIDA, ASAM

AMINO- ( Val,Pro,Pro ;

Ile,Pro,Pro)- Bbrp penelitian

A C E :1.Merpkan enzim yg

mengkatalisator Angiotensin Imenjadi Angiotensin II yangmerupakan senyawavasokontriktor kuat penyebabtekanan darah meningkat

2. Pengurai bradikinin(penyempitan pembuluhdarah)

A C E :1.Merpkan enzim yg

mengkatalisator Angiotensin Imenjadi Angiotensin II yangmerupakan senyawavasokontriktor kuat penyebabtekanan darah meningkat

2. Pengurai bradikinin(penyempitan pembuluhdarah)

FUNGSIONAL

KENAPA SUSUFERMENTASI??

ACEBahan

pertimbangan

- BIJI KEFIR……BAL- FUNGSI BAL- PEPTIDA, ASAM

AMINO- ( Val,Pro,Pro ;

Ile,Pro,Pro)- Bbrp penelitian

A C E :1.Merpkan enzim yg

mengkatalisator Angiotensin Imenjadi Angiotensin II yangmerupakan senyawavasokontriktor kuat penyebabtekanan darah meningkat

2. Pengurai bradikinin(penyempitan pembuluhdarah)

A C E :1.Merpkan enzim yg

mengkatalisator Angiotensin Imenjadi Angiotensin II yangmerupakan senyawavasokontriktor kuat penyebabtekanan darah meningkat

2. Pengurai bradikinin(penyempitan pembuluhdarah)

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FOTO GAMAL

Dgn ditemukanpenghambata ACE---susu fermentasi------hal sangat pentingpenghambatanhipertensi

ACE:1.Merpkan enzim yg

mengkatalisator Angiotensin Imenjadi Angiotensin II yangmerupakan senyawavasokontriktor kuat penyebabtekanan darah meningkat

2. Pengurai bradikinin(penyempitan pembuluh darah)

Dgn ditemukanpenghambata ACE---susu fermentasi------hal sangat pentingpenghambatanhipertensi

ACE sbgpertimbangan…….pengha

mbatan hipertensi

karena

Memegang perana pentingdalam mengontrol tekanan

darah

Memegang perana pentingdalam mengontrol tekanan

darah

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)

BERDASARKAN HAL INI DILAKUKANPENELITIAN MENGGUNAKAN SUSU FERMENTASIYAITU KEFIR (MENGANDUNG BAL DAN KHAMIR)

BALPROTEOLITIK…..PROTEIN.

DAYA SIMPAN SEDIKITINFORMASI

KARENA FAKTOR DAYA SIMPAN ERAT KAITANNYA DGN AKTIVITASMIKROORGANISME BIJI KEFIR (jumlahnya dan kualitas kimia)

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TUJUAN PENELITIAN 1. Diketahuinya kualitas kimia kefir konsumsi (protein,

lemak, laktosa, ph dan total asam ) yangmenghambat ACE selama penyimpanan

2. Diketahui umur simpan optimun dari kefirkonsumsi yang menghambat ACE tertinggi

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METODE PENELITIAN

PEUBAH:- TOTAL BAL- Kimia (pH, protein,lemak,laktosa,total asam)- Penghambatan ACE- IC50

PEUBAH:- TOTAL BAL- Kimia (pH, protein,lemak,laktosa,total asam)- Penghambatan ACE- IC50

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KEFIRKONSUMSI

KEFIRKONSUMSI

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HASIL

PerlakuanKefir konsumsi (hari)

0 3 6 9 12 SEM

Protein 4.75 a 4.46 bc 4.66 ab 4.26 cd 4.09 d 0.073

Tabel 1. Kualitas kimia kefir konsumsi selama penyimpananberbeda pada suhu 4 C

H

Protein 4.75 a 4.46 bc 4.66 ab 4.26 cd 4.09 d 0.073

Lemak 2.19 a 2.17 a 2.01 a 1.98 a 2.01 a 0.100

Laktosa 5.48 a 4.18 b 3.96 b 3.67 b 3.41 b 0.228

Total asam 1.97 d 2.56 c 2.68 bc 2.87 ab 2.95 a 0.046

pH 3.76 a 3.79 a 3.69 b 3.75 a 3.57 c 0.014

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PerlakuanKefir konsumsi (hari)

0 3 6 9 12 SEM

Total bakteriasam laktat(cfu/g)

2.39 x 10 8 4.35 x 10 7 1.92 x 10 8 8.63 x 10 7 2.82 x 10 6 3.9 x 10 7

Tabel 2. Total bakteri asam laktat dan E.coli kefir konsumsiselama penyimpanan berbeda pada suhu 4 C

Total bakteriasam laktat(cfu/g)

Total E.coli - - - - -

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Tabel 3. Penghambatan kefir konsumsi terhadap angiotensinconverting enzim selama penyimpanan pada suhu 4

Perlakuan

Kefir konsumsi (hari)

0 3 6 9 12

PenghambatanACE (%)

20.18 b 11.11 c 19.10 b 6.68 d 39.22 a

IC50 1.40 a 1.45 a 1.25 a 1.02 a 0.98 a

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KESIMPULAN

1. KEFIR KONSUMSI PADA MASA SIMPAN 0-12 MAMPUMENGHAMBAT ACE

2. KEFIR KONSUMSI PADA MASA SIMPAN 12 HARI MEMPUNYAIKEMAMPUAN TERTINGGI DALAM MENGHAMBAT ACESEBESAR 39,22 % DENGAN IC50 0,98 mg/ml

1. KEFIR KONSUMSI PADA MASA SIMPAN 0-12 MAMPUMENGHAMBAT ACE

2. KEFIR KONSUMSI PADA MASA SIMPAN 12 HARI MEMPUNYAIKEMAMPUAN TERTINGGI DALAM MENGHAMBAT ACESEBESAR 39,22 % DENGAN IC50 0,98 mg/ml

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TERIMAKASIH

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ANALISIS TOTAL MIKROBA

1mL

90 ml peptonwater 0,1%

10-1 10-2 10-7

METODE TUANG

Inkubasi padasuhu 28 0C selama

24 Jam

MEDIA :PLATE COUNT

AGAR (PCA)

90 ml peptonwater 0,1%

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ISOLASI BAKTERI BAL

…. . . ...

.

.

…. . . ...

.

.SAMPAI DIPEROLEHKOLONI HOMOGEN

Metode gores

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SELEKSI ISOLAT SECARA FENOTIP

UJI GRAM*Ambil isolat 1 ose*Kristal violet----1 menit*Cuci dgn air*Larutan iodine----1 menit*Alkohol 95 %*Cuci air*Safranin-----1 menit*Cuci ait*Keringkan --------mikroskop

*Ambil isolat 1 ose*Kristal violet----1 menit*Cuci dgn air*Larutan iodine----1 menit*Alkohol 95 %*Cuci air*Safranin-----1 menit*Cuci ait*Keringkan --------mikroskop

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Uji katalase• Isolat pada gelas obyek-----• ditetesi dgn 2 tetes hidrogen• peroksida 3 %

hasil positip------gelembonggas

• Isolat pada gelas obyek-----• ditetesi dgn 2 tetes hidrogen• peroksida 3 %

hasil positip------gelembonggas

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* UJI PRODUKSI GAS……homo danheterofermentatif

* UJI PERTUMBUHAN PADA SUHU 10, 37 DAN 45 C* UJI PERTUMBUHAN PADA Ph 2, 3, 4, 5,,6*uji sampai genus

* UJI PRODUKSI GAS……homo danheterofermentatif

* UJI PERTUMBUHAN PADA SUHU 10, 37 DAN 45 C* UJI PERTUMBUHAN PADA Ph 2, 3, 4, 5,,6*uji sampai genus

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• Uji Proteolitik……dengan cara:• 1. koloni yang sudah homogen/single• kultur, diambil dengan sulit gigi dan• ditumbuhkan pada media skin agar

• 2. sequensing

• Uji Proteolitik……dengan cara:• 1. koloni yang sudah homogen/single• kultur, diambil dengan sulit gigi dan• ditumbuhkan pada media skin agar

• 2. sequensing

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JAI /N6L.RENI ,INDAuAT

'EBAGAI PEiIAKALAH

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DINGAEDIAN (EPADA MA'YANAR I I'NlvENtlrII UDAVAT{AIEMBACA DENELITI II D iT