[ABSTRAK
Isolat Penicillium sp. ID10-T065 dimutasi menggunakan sinar Ultraviolet(UV), Etil Metil Sulfonat (EMS), dan kombinasi UV-EMS. Hasil mutasimenunjukkan bahwa aktivitas enzim β-glukosidase pada mutan lebih tinggidibandingkan wild-type (1,78 U/ml), kecuali pada mutan UM23. Mutan UV13mengalami peningkatan aktivitas β-glukosidase tertinggi (1,88 U/ml padaselobiosa 0,1% dan 5,53 U/ml pada selobiosa 1%), sedangkan mutan UM23menunjukkan aktivitas terendah (1,80 U/ml pada selobiosa 0,1% dan 1,75 U/mlpada selobiosa 1%). Aktivitas β-glukosidase pada mutan EM31 sebesar 1,86 U/mlpada selobiosa 0,1% dan 4,26 U/ml pada selobiosa 1%. Hasil analisis sekuen genβ-glukosidase 1 (bgl1) menunjukkan bahwa seluruh mutan mengalami mutasisubstitusi ketika dibandingkan dengan sekuen wild-type. Mutan UV13 mengalamiperubahan basa nukleotida paling banyak (7 basa) dibandingkan mutan EM31(5basa) dan UM23 (2 basa). Perubahan basa juga mengakibatkan gen mengalamimissense mutation sehingga terjadi kesalahan dalam penerjemahan kode asamamino, kecuali pada basa ke-2037 dari mutan UV13 dan basa ke-2034 serta 2037mutan EM31. Perubahan basa pada posisi tersebut tidak mengubah translasi asamamino (silent mutation). Hasil analisis sekuen gen bgl1 dan aktivitas enzimmenunjukkan bahwa sinar UV merupakan mutagen efektif untuk peningkatanaktivitas β-glukosidase pada isolat Penicillium sp. ID10-T065. Hasil identifikasisecara molekuler dan analisis pohon filogenetik, isolat Penicillium sp. ID10-T065memiliki kemiripan dan kekerabatan terdekat dengan spesies Penicilliumoxalicum.
ABSTRACT
The Penicillium sp. isolate ID10-T065 was mutated using Ultravioletirradiation (UV), Ethyl Methyl Sulfonate (EMS), and combination of UV -EMS.The results showed that β-glucosidase activity in the mutant was higher than thatof wild-type (1,78 U/ml), except for the mutant UM23. The β-glucosidase activityin mutant UV13 showed the highest activity (1,88 U/ml at cellobiose 0,1% and5,53 U/ml at cellobiose 1%), while mutant UM23 showed the lowest activity (1,80U/ml at cellobiose 0,1% and 1,75 U/ml at cellobiose 1%). The β-glucosidaseactivity of EM31 was 1,86 U/ml at cellobiose 0,1% and 4,26 U/ml at cellobiose1%. The results of the DNA sequence analysis of β-glucosidase 1 (bgl1) showedthat all mutants had substitution mutations when compared to the wild-typesequences. Mutant UV13 had the most base alteration (7 bases) compared to themutant EM31 (5 bases) and UM23 (2 bases). The bases alteration was leading tomissense mutation, except for the sequence of mutant UV13 at position 2037 andmutant EM31 at position 2034 and 2037. The base alteration of the sequence didnot change the amino acid translation (silent mutation). The results of the DNAsequences analysis of bgl1 and enzyme activities showed that UV light is aneffective mutagen to increase β-glucosidase activity in Penicillium sp. ID10-T065.The molecular identification and phylogenetic analysis showed that Penicilliumsp. ID10-T065 was closely related with Penicillium oxalicum., The Penicillium sp. isolate ID10-T065 was mutated using Ultravioletirradiation (UV), Ethyl Methyl Sulfonate (EMS), and combination of UV -EMS.The results showed that β-glucosidase activity in the mutant was higher than thatof wild-type (1,78 U/ml), except for the mutant UM23. The β-glucosidase activityin mutant UV13 showed the highest activity (1,88 U/ml at cellobiose 0,1% and5,53 U/ml at cellobiose 1%), while mutant UM23 showed the lowest activity (1,80U/ml at cellobiose 0,1% and 1,75 U/ml at cellobiose 1%). The β-glucosidaseactivity of EM31 was 1,86 U/ml at cellobiose 0,1% and 4,26 U/ml at cellobiose1%. The results of the DNA sequence analysis of β-glucosidase 1 (bgl1) showedthat all mutants had substitution mutations when compared to the wild-typesequences. Mutant UV13 had the most base alteration (7 bases) compared to themutant EM31 (5 bases) and UM23 (2 bases). The bases alteration was leading tomissense mutation, except for the sequence of mutant UV13 at position 2037 andmutant EM31 at position 2034 and 2037. The base alteration of the sequence didnot change the amino acid translation (silent mutation). The results of the DNAsequences analysis of bgl1 and enzyme activities showed that UV light is aneffective mutagen to increase β-glucosidase activity in Penicillium sp. ID10-T065.The molecular identification and phylogenetic analysis showed that Penicilliumsp. ID10-T065 was closely related with Penicillium oxalicum.] |
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