Two types of lipases (Lipases I and

Two types of lipases (Lipases I and II) were purified to homogeneity by Kohno et al.
(1994), using column chromatography on DEAE-Toyopearl. Lipase I consisted of two
polypeptide chains [a small peptide with sugar moiety (A-chain) and a large peptide of 34
kDa molecular weight (B-chain)]. Lipase II had a molecular mass of 30 kDa and a single
polypeptide chain (Kohno et al., 1994). Ohnishi et al. (1994b) reported an A. oryzae strain
that produced at least two kinds of extracellular lipolytic enzymes, L1 and L2. The enzyme
L1 was purified to homogeneity by ammonium sulfate and acetone fractionation, ion
exchange chromatography, and gel filtration. Lipase L1 was a monomeric protein (24 kDa
molecular weight) and preferentially cleaved all the ester bonds of triolein.
An extracellular lipase from Aci. calcoaceticus BD 413 was purified to homogeneity using
hydrophobic interaction fast performance liquid chromatography (FPLC) (Kok et al., 1995).
The enzyme had an apparent molecular mass of 32 kDa on SDS-PAGE and an optimal
activity pH of between 7.8 and 8.8 (Kok et al., 1995). Also, a lipase from Pe. roqueforti IAM
7268 was purified to homogeneity by a procedure involving ethanol precipitation, ammonium
sulfate precipitation, and three chromatographic steps on different matrices (DEAE-Toyopearl
650 M, Phenyl Toyopearl 650 M, Toyopearl HW-60). The molecular mass of purified lipase
was 25 kDa by electrophoresis (Mase et al., 1995). The enzyme had a high specificity
towards short-chain fatty acid esters (Mase et al., 1995). A Pichia burtonii lipase was purified
to homogeneity by a combination of DEAE-Sephadex A-50 ion exchange chromatography,
Sephadex G-100 gel filtration, and isoelectric focusing (Sugihara et al., 1995). The purified
enzyme was monomeric and had a molecular mass of 51 kDa by SDS-PAGE. The isoelectric
pH of the enzyme was 5.8 (Sugihara et al., 1995). The enzyme had temperature and pH
optima of 45
C and pH 6.5, respectively (Sugihara et al., 1995).
Kim et al. (1996) purified a highly alkaline extracellular lipase of Proteus vulgaris by ion
exchange chromatography. The purified lipase had a maximum hydrolytic activity at pH 10.0
and its molecular mass was 31 kDa by SDS-PAGE. Lin et al. (1996) purified an alkaline
lipase from P. pseudoalcaligenes F-111 to homogeneity. The apparent molecular mass by
SDS-PAGE was 32 kDa and the isoelectric pH was 7.3 (Lin et al., 1996). The enzyme
showed a preference for C
12
aryl and C
acyl groups when using p-nitrophenyl esters as
substrates. An extracellular lipase from P. aeruginosa KKA-5 was purified using ammonium
sulfate precipitation and successive chromatographic separations on hydroxyl appetite
(Sharon et al., 1998). After a 518-fold purification, the enzyme was homogenous electrophoretically
and its molecular mass was estimated to be 30 kDa (Sharon et al., 1998). The
enzyme was inhibited by SDS, an anionic surfactant; however, the cationic surfactants Triton
X-100 and Tween 80 appreciably enhanced the enzyme activity (Sharon et al., 1998).
14
A lipase produced by Staphylococcus epidermidis RP 62A was purified to homogeneity by
a combination of precipitation techniques, metal affinity chromatography, and gel filtration
(Simons et al., 1998). The purified enzyme had a pH optimum of 6.0 and required calcium as
a cofactor for catalytic activity (Simons et al., 1998). A recombinant lipase (rROL) produced
by S. cerevisiae was purified by ethanol precipitation, butyl-Toyopearl 650 M chromatography,
and Sephacryl S-100 HR gel filtration, to a single band by native PAGE (Takahashi
et al., 1998). The band was found to consist of two proteins with molecular masses of 35 and
46 kDa, on SDS-PAGE.

Hasil (Bahasa Indonesia) 1:

Two types of lipases (Lipases I and II) were purified to homogeneity by Kohno et al.(1994), using column chromatography on DEAE-Toyopearl. Lipase I consisted of twopolypeptide chains [a small peptide with sugar moiety (A-chain) and a large peptide of 34kDa molecular weight (B-chain)]. Lipase II had a molecular mass of 30 kDa and a singlepolypeptide chain (Kohno et al., 1994). Ohnishi et al. (1994b) reported an A. oryzae strainthat produced at least two kinds of extracellular lipolytic enzymes, L1 and L2. The enzymeL1 was purified to homogeneity by ammonium sulfate and acetone fractionation, ionexchange chromatography, and gel filtration. Lipase L1 was a monomeric protein (24 kDamolecular weight) and preferentially cleaved all the ester bonds of triolein.An extracellular lipase from Aci. calcoaceticus BD 413 was purified to homogeneity usinghydrophobic interaction fast performance liquid chromatography (FPLC) (Kok et al., 1995).The enzyme had an apparent molecular mass of 32 kDa on SDS-PAGE and an optimalactivity pH of between 7.8 and 8.8 (Kok et al., 1995). Also, a lipase from Pe. roqueforti IAM7268 was purified to homogeneity by a procedure involving ethanol precipitation, ammoniumsulfate precipitation, and three chromatographic steps on different matrices (DEAE-Toyopearl650 M, Phenyl Toyopearl 650 M, Toyopearl HW-60). The molecular mass of purified lipasewas 25 kDa by electrophoresis (Mase et al., 1995). The enzyme had a high specificitytowards short-chain fatty acid esters (Mase et al., 1995). A Pichia burtonii lipase was purifiedto homogeneity by a combination of DEAE-Sephadex A-50 ion exchange chromatography,Sephadex G-100 gel filtration, and isoelectric focusing (Sugihara et al., 1995). The purifiedenzyme was monomeric and had a molecular mass of 51 kDa by SDS-PAGE. The isoelectricpH of the enzyme was 5.8 (Sugihara et al., 1995). The enzyme had temperature and pHoptima of 45
C and pH 6.5, respectively (Sugihara et al., 1995).Kim et al. (1996) purified a highly alkaline extracellular lipase of Proteus vulgaris by ionexchange chromatography. The purified lipase had a maximum hydrolytic activity at pH 10.0and its molecular mass was 31 kDa by SDS-PAGE. Lin et al. (1996) purified an alkalinelipase from P. pseudoalcaligenes F-111 to homogeneity. The apparent molecular mass bySDS-PAGE was 32 kDa and the isoelectric pH was 7.3 (Lin et al., 1996). The enzymeshowed a preference for C12aryl and Cacyl groups when using p-nitrophenyl esters assubstrates. An extracellular lipase from P. aeruginosa KKA-5 was purified using ammoniumsulfate precipitation and successive chromatographic separations on hydroxyl appetite(Sharon et al., 1998). After a 518-fold purification, the enzyme was homogenous electrophoreticallyand its molecular mass was estimated to be 30 kDa (Sharon et al., 1998). Theenzyme was inhibited by SDS, an anionic surfactant; however, the cationic surfactants TritonX-100 dan Tween 80 lumayan meningkatkan aktivitas enzim (Sharon et al., 1998).14Lipase yang diproduksi oleh Staphylococcus epidermidis RP 62A disucikan untuk keseragaman olehkombinasi dari teknik curah hujan, logam afinitas kromatografi dan gel filtrasi(Simons et al. 1998). Enzim murni memiliki pH optimal kalsium 6.0 dan diperlukan sebagaikofaktor untuk aktivitas (Simons et al., 1998). Lipase rekombinan (rROL) yang diproduksioleh S. cerevisiae disucikan oleh etanol curah hujan, butil-Toyopearl 650 M kromatografi,dan Sephacryl S-100 HR gel filtrasi, untuk sebuah band yang satu laman asli (Takahashiet al. 1998). Band ditemukan terdiri dari kedua protein dengan massa molekul 35 dankDa 46, di SDS-halaman.