Lipase Enzymes Purification from the Chamomile (Matricaria chamomilla L.) and Its Immobilization on Various Supports

Year 2018, Volume: 8 Issue: 1, 131 - 143, 18.06.2018

Bahar Bilgin Sökmen , Burçak Sarı Tuğba Azap

https://doi.org/10.31466/kfbd.396563

Abstract

In this study, lipase enzyme was firstly purified from chamomile (Matricaria chamomilla L.) which is abundant in
Giresun (Turkey) and surroundings and the kinetic properties of the enzyme were investigated. 70 % of (NH4)2SO4
fraction of lipase esterase activity homogenate was obtained. After dialysis, this fraction was applied to DEAE-cellulose
column, and lipase was purified 26 times. In the purification process, the protein amount was determined by using
Lowry and E280/E260 Warburg methods while lipase esterase activtiy was assayed with Erlanson’s method. It was found
that the purified enzyme, which obtained with SDS-PAGE electroforesis, had a molecular weight of 30 kDa. The
optimum pH and temperature values were determined. It was found that the lipase purified from chamomile had an
optimum pH value of 9 and an optimum temperature of 50 °C. When the affinity against different substrates of the
enzyme was examined, the highest value was determined in p-nitrophenyl palmitate, with respectively Km and Vmax
values being 0.2899 mM and 144.93 Units. Also, the lipase enzyme purified from chamomile was immobilized on
various carriers by adsorption, covalent and ionic binding methods. The kinetic properties of the immobilized
chamomile lipase such as optimum pH, optimum temperature were also studied.

Keywords

Chamomile (Matricaria chamomilla L.), Lipase, Purification, Characterization

Papatyadan (Matricaria chamomilla L.) Lipaz Enziminin Saflaştırılması ve Çeşitli Taşıyıcılara İmmobilize Edilmesi

Abstract

Bu çalışmada, Giresun İli ve çevresinde bol miktarda
yetişen papatyadan (Matricaria chamomilla
L.) lipaz enzimi ilk defa
saflaştırıldı ve kinetik özellikleri incelendi. Lipaz
esteraz aktivitesi gösteren homojenizatın % 70’lik (NH₄)₂SO₄
kesiti elde edildi. Dializden sonra bu kesit DEAE-selüloz kolonlara uygulanarak
lipaz enzimi 26 kat saflaştırıldı. Saflaştırma işlemlerinde protein miktarı
Lowry ve E₂₈₀/E₂₆₀ Warburg yöntemlerine göre, lipaz
esteraz aktivitesi ise Erlanson yöntemine göre tayin edildi. SDS-PAGE
elektroforezi ile elde edilen saflaştırılmış enzimin molekül ağırlığının 30 kDa
olduğu bulundu. Optimum pH ve sıcaklık değerleri belirlendi. Papatyadan
saflaştırılan lipazın optimum pH’sının 9, optimum sıcaklığının 50 °C olduğu
bulundu. Enzimin çeşitli substratlara karşı ilgisi incelendiğinde, lipazın
ilgisinin en çok p-nitrofenil palmitatta olduğu ve bu substrata karşı K_m
ve V_max değerlerinin sırasıyla 0,2899 mM ve 144,93 Ünite olduğu
saptandı.Ayrıca, papatyadan saflaştırılan lipaz enzimi çeşitli
taşıyıcılar  üzerine
adsorpsiyon, kovalent ve iyonik bağlama yöntemleri ile immobilize edildi. İmmobilize
edilmiş papatya lipazının optimum pH, optimum sıcaklık gibi kinetik özellikleri
de incelendi.

Keywords

Papatya (Matricaria chamomilla L.), Lipaz, Saflaştırma, İmmobilizasyon

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