product is formed. The absorbance o

product is formed. The absorbance of the samples at 405 nm can
be measured with a microtiter plate (ELISA) reader and is directly
correlated to the level of RT activity. A fixed amount (4–6 ng) of
recombinant HIV-1 RT was used. The inhibitory activity of the
lectin was calculated as percent inhibition as compared to a
control without the protein (Wang and Ng, 2001). The mushroom
ribosome-inactivating protein hypsin was used as a positive
control (Lam and Ng, 2001).
Assay for ribonuclease activity
The assay was performed because some lectins display this
activity (Liu et al., 2007). Ribonuclease activity of RLL toward
yeast tRNA (Sigma) was assayed by determining the generation of
acid-soluble, UV absorbing species with the method of Mocket al.
(Mock et al., 1996). The lectin was incubated with 200mg of tRNA
in 150mg 100 mM morpholino-ethanesulfonic acid buffer (pH
6.0) at 371C for 1 h. The reaction was terminated by adding 350ml
of ice-cold 3.4% perchloric acid. After leaving on ice for 15 min, the
sample was centrifuged (15,000g, 15 min) at 41C. The OD 260
of the supernatant was read after appropriate dilution. One unit of
enzymatic activity was defined as the amount of enzyme that
brings about an increase of one per minute in OD 260 nm in the
acid-soluble supernatant (Mock et al., 1996). Bovine pancreatic
RNase A (Sigma) was used as a positive control.
Assay for antifungal activity
The assay was carried out since some lectins demonstrate this
activity (Ciopraga et al., 1999). The assay for antifungal activity
toward Fusarium oxysporum, Rhizoctonia cerealis, Rhizoctonia
solani, and Sclerotinia sclerotiorum was carried out in
10015 mm petri dishes containing 10 ml of potato dextrose
agar (PDA). After development of the mycelial colony, sterile
blank paper disks (0.625 cm in diameter) were laid at a distance of
0.5 cm away from the rim of the mycelial colony. An aliquot
(15ml) of the lectin was added to a disk. The dishes were
incubated at 231C for 72 h until mycelial growth had enveloped
the disks containing the control and had produced crescents of
inhibition around disks containing samples with antifungal
activity (Wang and Ng, 2006; Ye et al., 2001). The mushroom
ribosome-inactivating protein hypsin was used as a positive
control (Lam and Ng, 2001).
Results and discussion
Purification of RLL
The fraction of the fruiting body extract unadsorbed on DEAEcellulose (D1) but not the adsorbed fractions (D2,3,4) contained the
hemagglutinating activity (Table 1). Fraction D1 was divided into

Hasil (Bahasa Indonesia) 1:

Produk ini terbentuk. Absorbansi sampel di 405 nm dapatdiukur dengan pembaca piring (ELISA) Sungkup asap dan langsungberkorelasi untuk tingkat RT aktivitas. Tetap sebesar (4 – 6 ng)rekombinan RT HIV-1 digunakan. Aktivitas penghambatanlektin dihitung sebagai inhibisi persen dibandingkan denganmengendalikan tanpa protein (Wang dan Ng, 2001). Jamurinactivating ribosom protein hypsin digunakan sebagai positifkontrol (Lam dan Ng, 2001).Assay untuk aktivitas katalisisAssay dilakukan karena lectins beberapa tampilan iniaktivitas (Liu et al., 2007). Aktivitas katalisis RLL menujuragi tRNA (Sigma) diuji dengan menentukan generasiasam larut, UV menyerap jenis dengan metode Mocket al.(Pura-Pura et al., 1996). Lektin diinkubasi dengan 200mg tRNAdalam 150mg 100 mM morpholino-ethanesulfonic buffer asam (pH6.0) pada 371 C untuk 1 h. Reaksi diakhiri dengan menambahkan 350mldingin 3,4% asam perklorat pekat. Setelah meninggalkan di atas es untuk 15 menitsampel disentrifugasi (15.000 g, 15menit) pada 41C. OD 260dari supernatant dibaca setelah diencerkan. Satu unitaktivitas enzim didefinisikan sebagai jumlah enzim yangmembawa peningkatan satu per menit di dari 260 nm diasam larut supernatant (Mock et al., 1996). Pankreas sapiRNase (Sigma) digunakan sebagai kontrol positif.Assay untuk aktivitas anti jamurAssay dilakukan karena lectins beberapa menunjukkan hal iniaktivitas (Ciopraga et al., 1999). Assay aktivitas anti jamurmenuju Fusarium oxysporum, pelepah Rhizoctonia selepada, pelepah Rhizoctoniasolani, dan Sclerotinia sclerotiorum dilakukan di100 15 mm piring petri berisi 10 ml dekstrosa kentangagar-agar (PDA). Setelah pembangunan koloni mycelial, sterilkertas kosong disk (0,625 cm diameter) diletakkan pada jarak0,5 cm dari tepi mycelial koloni. Aliquot(15ml) dari lektin ditambahkan ke disk. Piring yangdiinkubasi 231 c untuk 72 h sampai mycelial pertumbuhan telah ditutupidisk berisi kontrol dan telah menghasilkan Crescent dariinhibisi di sekitar disk yang berisi sampel dengan antijamuraktivitas (Wang dan Ng, 2006; Ye et al., 2001). Jamurinactivating ribosom protein hypsin digunakan sebagai positifkontrol (Lam dan Ng, 2001).Hasil dan diskusiPemurnian RLLSebagian kecil dari tubuh berbuah ekstrak unadsorbed pada DEAEcellulose (D1) tetapi tidak adsorbed fraksi (D2, 3, 4) terdapathemagglutinating kegiatan (Tabel 1). Fraksi D1 dibagi menjadi