Aprilia Yean Wisaka

161431005

2 Analis Kimia

KUIS BIOTEKNOLOGI

GENBANK : AB025721.1

A. Identitas Sekuen
1. Definition : Thermococcus sp. B1001 cgtA gene for cyclodextrin
glucanotransferase, completete cds.
2. Organism : Thermococcus sp. B1001
Archae; Euryarchaeota; Thermococci; Thermococcales;
Thermococcaceae; Thermococcus.
3. Mol type : Genomic DNA
4. Gene : cgtA (55 – 2274)
5. Regulatory : Ribosom bindeng site (42 – 53)
6. CDS : 55 – 2274
7. Product : Cyclodextrin glucanotransferase
8. Protein ID : BAA88217.1
9. Sig. Peptide : 55 - 135
10. Origin :
1 tatcattaat agtgatacat tattaatagt gatacatttt tggagggaga tgacatgaga
61 agcagaaaga tttatatgct tgttgtattg cttctttttt taggatttag tgaacaaatt
121 tcaacagtag ctgcaggggt cagtccctca tatccagcag gggatcccca gacatgggtt
181 atctaccaaa tagtcattga tagattttac gatgggaaca cctcaaacaa tgatccatta
241 aaaagtccag gattatatga tccaaacaaa gagaactgga agctttattg gggcggtgat
301 ttagagggca tcatagcaaa gctgccctac ttatatgaac ttggagtgag tgccatatgg
361 atttctccag tgtttgataa tatagatgtt cctatcaatg gaagcaatgg tctagaggcg
421 gggtatcatg ggtattggcc aaaggacttt aaggttatag aagagcactt tgggacatgg
481 gaaatattcc gcagacttag tcaagaagca gccaagtaca atatcacgat tataattgac
541 ttcgttccaa accacagcaa tcccaatgat gcgggcgagt atggagctct atatgataat
601 ggcacgtttg ttattgatta tccaacggat gcaaattatg cgactgttca cccaataaca
661 aagagtttat cgtacatata caatcacaat ggaggaataa caaattggaa tgatagatgg
721 gaagtcagat ataaaaatct ctttaatctg gctgatctta atcagttgaa tccatgggtg
781 gacaattatc ttaaagaatc gacagtatct tacttagaag ctggaattgg aggaataaga
841 atagatgcag ttaaacactt agagcctgga tggctcaaaa cttatgcaga ctatgtatat
901 gcaagaaaga acgtatttat gtttggagaa tggtaccagg gctttaacga tgagatgtac
961 tgggatatgg ttaagtttgc aaactatacg gggataggcc ttattaacat cccactccaa
1021 caagttctcg ttgatgtttt tgcttatgat acaaagacta tgtgggactt ggaaagcgct
1081 gttaataaat ataccattga ctttatgtgg caaaataaac taactatatt cgtagacagc
1141 cacgatgttc caagatttct gtccttgaga aatgacttaa taagatttca ccaagctctt
1201 gcttttgttc taaccgctcc tggaattcca ataatctatt acggtgatga acaatatctt
1261 cacaatgata ctgtaaatga ctttggtcaa gttggaggtg acccatataa tagacctatg
1321 atgaaaactt ggaacacatc aacaaccgct tttaaactaa taaagactct cgcaagtctg
1381 agacgctata atcctgcttt agcttatgga ttgatcgcga caaggtatgt aagcgatgac
1441 atttacatct ttgaaagaaa gttctttgat aacgttgttc tcgtagcaat aaatcgcaat
1501 ttaaattctc catatgtggt ctctaacgtt tacacctccc taccagatgg aagttacaat
1561 gactaccttg gaggactttt gaacggagtt ggaataacgg tttcatctgg tacattctct
1621 gtagagttac ctgcaggttc agtatcagta tggcagtaca aagcaacgcc gactgatcca
1681 tgggtaggag caattgatcc agttatggga agagctggaa acattgtcac agttagtgga
1741 gaagggtttg gggatgttcc gggaagagtc ttaataacaa atggacaaga ttactggact
1801 gcagaagtta catattggag cgataaaagc gtagagttca tagttccttc aggaataact
1861 acccaactaa acgaaaatca tgttgaagtc agaattgaaa gggctgatgg ggctacatca
1921 aatggaatag cctttgaata tctaacgaac aaacaaattc cagctatttt tgaagttagg
1981 aacactcaag gtacaaattt agaaacccaa gttggagaat ttttatggct aacaggtagt
2041 gttccagagt tgagctattg gagtccagaa acaataaaag ctgttggacc aatgttgtgt
2101 ccaggatggc ctgattggtt tgttgtagca agtgtcccgg cggataccta tatagagttc
2161 aagttcttaa aggcaccgct tggaggcact ggaatctggg aagtaggaag caatcacgca
2221 tacctaacac cttcaagcgg aataggggaa gtttcagttg aagccaaccg ttgatattct
2281 tttttattat tcttattgca tctatttgac attattgtat tgtttagttc atatgtcaat
2341 tcgtgagcaa tgaagcgtaa ggtatatata tcactg//

11. Reference
1
AUTHORS : Yamamoto,T., Shiraki,K., Fujiwara,S., Takagi,M.,
Fukui,K. And Imanaka,T.

TITLE : In vitro heat effect on functional and conformational

changes of cyclodextrin glucanotransferase from

hyperthermophilic archaea

JOURNAL : Biochem. Biophys. Res. Commun. 265 (1), 57-61

(1999)

PUBMED : 10548490

2 (bases 1 to 2376)

AUTHORS : Fujiwara,S., Yamamoto,T. and Imanaka,T.

TITLE : Direct Submission

JOURNAL : Submitted (02-APR-1999) Shinsuke Fujiwara, Osaka

University, Department of Biotecnology; 2-1

Yamadaoka, Suita, Osaka 565-0871, Japan (E-

mail:s.fujiwara@cell.bio.eng.osaka-u.ac.jp, Tel:81-6-

877-5111(ex.3445), Fax:81-6-879-7441
B. FASTA

>AB025721.1 Thermococcus sp. B1001 cgtA gene for cyclodextrin glucanotransferase,

complete cds
TATCATTAATAGTGATACATTATTAATAGTGATACATTTTTGGAGGGAGATGACATGAGAAGCAGAAAGA
TTTATATGCTTGTTGTATTGCTTCTTTTTTTAGGATTTAGTGAACAAATTTCAACAGTAGCTGCAGGGGT
CAGTCCCTCATATCCAGCAGGGGATCCCCAGACATGGGTTATCTACCAAATAGTCATTGATAGATTTTAC
GATGGGAACACCTCAAACAATGATCCATTAAAAAGTCCAGGATTATATGATCCAAACAAAGAGAACTGGA
AGCTTTATTGGGGCGGTGATTTAGAGGGCATCATAGCAAAGCTGCCCTACTTATATGAACTTGGAGTGAG
TGCCATATGGATTTCTCCAGTGTTTGATAATATAGATGTTCCTATCAATGGAAGCAATGGTCTAGAGGCG
GGGTATCATGGGTATTGGCCAAAGGACTTTAAGGTTATAGAAGAGCACTTTGGGACATGGGAAATATTCC
GCAGACTTAGTCAAGAAGCAGCCAAGTACAATATCACGATTATAATTGACTTCGTTCCAAACCACAGCAA
TCCCAATGATGCGGGCGAGTATGGAGCTCTATATGATAATGGCACGTTTGTTATTGATTATCCAACGGAT
GCAAATTATGCGACTGTTCACCCAATAACAAAGAGTTTATCGTACATATACAATCACAATGGAGGAATAA
CAAATTGGAATGATAGATGGGAAGTCAGATATAAAAATCTCTTTAATCTGGCTGATCTTAATCAGTTGAA
TCCATGGGTGGACAATTATCTTAAAGAATCGACAGTATCTTACTTAGAAGCTGGAATTGGAGGAATAAGA
ATAGATGCAGTTAAACACTTAGAGCCTGGATGGCTCAAAACTTATGCAGACTATGTATATGCAAGAAAGA
ACGTATTTATGTTTGGAGAATGGTACCAGGGCTTTAACGATGAGATGTACTGGGATATGGTTAAGTTTGC
AAACTATACGGGGATAGGCCTTATTAACATCCCACTCCAACAAGTTCTCGTTGATGTTTTTGCTTATGAT
ACAAAGACTATGTGGGACTTGGAAAGCGCTGTTAATAAATATACCATTGACTTTATGTGGCAAAATAAAC
TAACTATATTCGTAGACAGCCACGATGTTCCAAGATTTCTGTCCTTGAGAAATGACTTAATAAGATTTCA
CCAAGCTCTTGCTTTTGTTCTAACCGCTCCTGGAATTCCAATAATCTATTACGGTGATGAACAATATCTT
CACAATGATACTGTAAATGACTTTGGTCAAGTTGGAGGTGACCCATATAATAGACCTATGATGAAAACTT
GGAACACATCAACAACCGCTTTTAAACTAATAAAGACTCTCGCAAGTCTGAGACGCTATAATCCTGCTTT
AGCTTATGGATTGATCGCGACAAGGTATGTAAGCGATGACATTTACATCTTTGAAAGAAAGTTCTTTGAT
AACGTTGTTCTCGTAGCAATAAATCGCAATTTAAATTCTCCATATGTGGTCTCTAACGTTTACACCTCCC
TACCAGATGGAAGTTACAATGACTACCTTGGAGGACTTTTGAACGGAGTTGGAATAACGGTTTCATCTGG
TACATTCTCTGTAGAGTTACCTGCAGGTTCAGTATCAGTATGGCAGTACAAAGCAACGCCGACTGATCCA
TGGGTAGGAGCAATTGATCCAGTTATGGGAAGAGCTGGAAACATTGTCACAGTTAGTGGAGAAGGGTTTG
GGGATGTTCCGGGAAGAGTCTTAATAACAAATGGACAAGATTACTGGACTGCAGAAGTTACATATTGGAG
CGATAAAAGCGTAGAGTTCATAGTTCCTTCAGGAATAACTACCCAACTAAACGAAAATCATGTTGAAGTC
AGAATTGAAAGGGCTGATGGGGCTACATCAAATGGAATAGCCTTTGAATATCTAACGAACAAACAAATTC
CAGCTATTTTTGAAGTTAGGAACACTCAAGGTACAAATTTAGAAACCCAAGTTGGAGAATTTTTATGGCT
AACAGGTAGTGTTCCAGAGTTGAGCTATTGGAGTCCAGAAACAATAAAAGCTGTTGGACCAATGTTGTGT
CCAGGATGGCCTGATTGGTTTGTTGTAGCAAGTGTCCCGGCGGATACCTATATAGAGTTCAAGTTCTTAA
AGGCACCGCTTGGAGGCACTGGAATCTGGGAAGTAGGAAGCAATCACGCATACCTAACACCTTCAAGCGG
AATAGGGGAAGTTTCAGTTGAAGCCAACCGTTGATATTCTTTTTTATTATTCTTATTGCATCTATTTGAC
ATTATTGTATTGTTTAGTTCATATGTCAATTCGTGAGCAATGAAGCGTAAGGTATATATATCACTG

C. Translasi
1. Hasil translasi :
"MRSRKIYMLVVLLLFLGFSEQISTVAAGVSPSYPAGDPQTWVIYQIVIDRFYDGNTSNNDPLK
SPGLYDPNKENWKLYWGGDLEGIIAKLPYLYELGVSAIWISPVFDNIDVPINGSNGLEAGYHGY
WPKDFKVIEEHFGTWEIFRRLSQEAAKYNITIIIDFVPNHSNPNDAGEYGALYDNGTFVIDYPT
DANYATVHPITKSLSYIYNHNGGITNWNDRWEVRYKNLFNLADLNQLNPWVDNYLKESTVSYLE
AGIGGIRIDAVKHLEPGWLKTYADYVYARKNVFMFGEWYQGFNDEMYWDMVKFANYTGIGLINI
 PLQQVLVDVFAYDTKTMWDLESAVNKYTIDFMWQNKLTIFVDSHDVPRFLSLRNDLIRFHQALA
FVLTAPGIPIIYYGDEQYLHNDTVNDFGQVGGDPYNRPMMKTWNTSTTAFKLIKTLASLRRYNP
ALAYGLIATRYVSDDIYIFERKFFDNVVLVAINRNLNSPYVVSNVYTSLPDGSYNDYLGGLLNG
VGITVSSGTFSVELPAGSVSVWQYKATPTDPWVGAIDPVMGRAGNIVTVSGEGFGDVPGRVLIT
NGQDYWTAEVTYWSDKSVEFIVPSGITTQLNENHVEVRIERADGATSNGIAFEYLTNKQIPAIF
EVRNTQGTNLETQVGEFLWLTGSVPELSYWSPETIKAVGPMLCPGWPDWFVVASVPADTYIEFK
FLKAPLGGTGIWEVGSNHAYLTPSSGIGEVSVEANR"

2. Ukuran sekuen nukleotida : 2376 bp

D. Protein Cystein
1. Jumlah protein cystein yang terdapat pada sekuen : 1 (satu)
2. Kemungkinan terbentuk jembatan disulfida : Tidak ada,
karena hanya
terdapat satu
protein cystein

E. Fungsi Cyclodextrin Glucanotransferase

Cyclodextrin glucanotransferase or CGTase is a bacterial enzyme belonging

to the same family of the α-amylase specifically known as glycosyl-hydrolase family
13. This peculiar enzyme is capable of catalyzing more than one reaction with the most
important being the synthesis of non-reducing cyclic dextrins known as cyclodextrins
starting from starch, amylose, and other polysaccharides. CGTase is an enzyme
common to many bacterial species, in particular of the Bacillus genus (e.g. B. circulans,
B. macerans and B. stearothermophilus), as well as to some archaea, but it is not known
to be present in any other species.

Catalytic activities

All of the CGTases can catalyze up to four reactions: cyclization, coupling,

disproportionation and hydrolysis. All these activities share the same catalytic
mechanism which is common to all glycosyl-hydrolases.

Cyclization is the process through which a linear polysaccharidic chain is

cleaved and the two ends of the cleaved fragment are joined to produce a circular
dextrin (cyclodextrin or CD): on the basis of the number of sugar residues this circular
product is made of three main type of cyclodextrins can be distinguished, α-CD with six
residues, β-CD with seven residues and γ-CD with eight residues.
 The coupling reaction can be easily described as the reverse process of
cyclization: the enzyme cleaves a cyclodextrin to produce a linear dextrin which is
subsequently joined to a linear oligosaccharide.

Disproportionation is very similar to coupling, but the cleaved dextrin is not a

cyclodextrin, but a linear oligosaccharide that is then joined to a second oligosaccharide.

CGTase also has a weak hydrolyzing activity which consists in cleaving the
longer polysaccharidic chains into shorter fragments. (Sumber : Wikipedia)

Dari urutan sekuen protein berikut ini :

SAYAMAHASISWAANALISKIMIAPOLITEKNIKNEGERIBANDUNGYAN
GBERALAMATDICIWARUGAKECAMATANPARONGPONGKABUPATE
NBANDUNGBARAT

- Sebutkan nama asam amino dari sekuen di atas.

- Adakah sekuen yang bukan asam amino? Jika ada sebutkan, dan ada berapa
banyaknya?

Jawab :

1. Nama Asam Amino dari Sekuen

Serine-Alanine-Tyrosine-Alanine-Methionine-Alanine-Histidine-Alanine-
Serine-Isoleucine-Serine-Typtophan-Alanine-Alanine-Asparagine-Alanine-
Leucine-Isoleucin-Serine-Lysine-Isoleucine-Methionine-Isoleucine-Alanine-
Prolin-O-Leucine-Isoleucine-Threonine-Glutamic Acid-Lysine-Asparagine-
Isoleucine-Lysine-Asparagine-GlutamicAcid-Glycine-GlutamicAcid
Arginine-Isoleucine-B-Alanine-Asparagine-AsparticAcid-U-Asparagine-
Glycine-Tyrosine-Alanine-Asparagine-Glycine-B-GlutamicAcid-Arginine-
Alanine-Leucin-Alanine-Methionine-Alanine-Threonine-AsparticAcid-
Isoleucine-Cysteine-Isoleucine-Tryptophan-Alanine-Arginine-U-Glycine-
Alanine-Lysine-GlutamicAcid-Cysteine-Alanine-Methionine-Alanine-
Threonine-Alanine-Asparagine-Prolin-Alanine-Arginine-O-Asparagine-
Glycine-Prolin-O-Asparagine-Glycine-Lysine-Alanine-B-U-Prolin-Alanine-
Threonine-GlutamicAcid-Asparagin-B-Alanine-Asparagine-AsparticAcid-
U-Asparagine-Glycine-B-Alanine-Arginine-Alanine-Threonine

S = Serine
A = Alanine
Y = Tyrosine
M = Methionine
 H = Histidine
I = Isoleucine
W = Tryptophan
N = Asparagine
L = Leucine
K = Lysine
P = Prolin
T = Threonine
E = Glutamic Acid
G = Glycine
R = Arginine
D = Aspartic Acid
C = Cysteine

2. Yang bukan asam amino ada 3 jenis, yaitu O (ada 3 buah), B (ada 5 buah),
dan U (ada 4 buah).