{"id":400,"date":"2025-03-10T16:05:56","date_gmt":"2025-03-10T09:05:56","guid":{"rendered":"https:\/\/dnape.online\/?p=400"},"modified":"2025-03-21T14:47:27","modified_gmt":"2025-03-21T07:47:27","slug":"pse31-i","status":"publish","type":"post","link":"https:\/\/dnape.online\/en\/2025\/03\/pse31-i\/","title":{"rendered":"Restriction Endonuclease Pse31I from Bacterial Strain Peribacillus Species 31, an Isoschizomer of Eco31I, Recognizes the Non-palindromic DNA Sequence 5\u2019-GGTCTC-3\u2019 (1\/5)"},"content":{"rendered":"\n<\/span>

[vc_row css=”.vc_custom_1576496196814{background-color: #ededed !important;}”][vc_column width=”2\/3″][vc_column_text]V.A. Chernukhin1<\/sup>, V.S. Dedkov1<\/sup>, N.A. Mikhnenskova1<\/sup>, N.A. Vorontsova1<\/sup>, M.A. Abdurashitov1<\/sup>, D.A. Gonchar1<\/sup>, S.Kh. Degtyarev<\/a>1<\/sup><\/strong><\/p>\n

1<\/sup> SibEnzyme LLC, Novosibirsk, Russia<\/p>\n

* Corresponding author: Valeriy Chernukhin, SibEnzyme LLC, 2\/12 Timakova St., Novosibirsk, 630117, Russia; Tel.: +7(383)333-4991; Fax: +7(383)333-6853; E-mail: valerunishe@mail.ru<\/a><\/p>\n

\n

We have discovered a bacterial strain, Peribacillus species<\/em> 31, which produces a novel restriction endonuclease named Pse31I. The enzyme recognizes the non-palindromic hexanucleotide DNA sequence 5\u2019-GGTCTC-3\u2019 and cleaves outside the recognition site at positions 1\/5: 5\u2019-GGTCTC(N)1<\/sub>^ \/ 3\u2019-CCAGAG(N)5<\/sub>^ -5\u2019. Thus, Pse31I is a true isoschizomer of the restriction enzyme Eco31I [1]. The producer strain was identified by morphological and biochemical characteristics as well as by analysis of the 16S rRNA gene fragment.
\nPse31I shows maximum activity at 37\u00b0C and is inactivated by incubation at 55 and 65\u00b0C. The enzyme does not cleave sites with methylated cytosine in the top strand (5\u2019-GGTCT(5mC)-3\u2019) and\/or in the bottom strand (3\u2019-(5mC)CAGAG).<\/p>\n

The optimal conditions for enzyme activity are SE-buffer 5 (Y) (33 mM Tris-acetate (pH 7.9), 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT) at 37\u00b0C.<\/p>\n

A preparation of the restriction endonuclease PecI with a concentration of 10,000 U\/mL was obtained through a four-step chromatographic purification. The optimal reaction conditions are SE-buffer 5 (Y) (33 mM Tris-acetate (pH 7.9), 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT) and a temperature of 37o<\/sup>C.<\/p>\n

Abbreviations:<\/em> BSA \u2013 bovine serum albumin; DTT \u2013 dithiothreitol; T7 DNA \u2013 bacteriophage T7 DNA; \u03bb DNA \u2013 bacteriophage \u03bb DNA; Tris \u2013 tris(hydroxymethyl)aminomethane; PAGE \u2013 polyacrylamide gel; restriction enzyme, RE \u2013 restriction endonuclease; EDTA \u2013 ethylenediaminetetraacetic acid; SAM \u2013 S-adenosyl-L-methionine; M.SssI \u2013 SssI DNA methyltransferase; (5mC) \u2013 5-methylcytosine;<\/p>\n<\/div>\n

[\/vc_column_text][\/vc_column][vc_column width=”1\/3″][vc_column_text]Keywords:<\/strong> producer strain, enzyme isolation, type IIS restriction endonuclease, sensitivity to (5mC)G DNA methylation<\/p>\n

DOI: <\/strong>10.26213\/3034-4301.2025.4.1.001<\/a><\/p>\n

Citation:<\/strong><\/p>\n

V.A. Chernukhin, V.S. Dedkov, N.A. Mikhnenskova, N.A. Vorontsova, M.A. Abdurashitov, D.A. Gonchar, S.Kh. Degtyarev. (2025) Restriction Endonuclease Pse31I from Bacterial Strain Peribacillus Species 31, an Isoschizomer of Eco31I, Recognizes the Non-palindromic DNA Sequence 5\u2019-GGTCTC-3\u2019 (1\/5), DNA-Processing Enzymes, vol. 2025(1), DOI: 10.26213\/3034-4301.2025.4.1.001<\/p>\n

\"Creative<\/a>
\nThis article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License<\/a>[\/vc_column_text][\/vc_column][\/vc_row]<\/p>","protected":false},"excerpt":{"rendered":"

We have discovered a bacterial strain, Peribacillus species 31, which produces a novel restriction endonuclease named Pse31I. The enzyme recognizes a six-nucleotide non-palindromic DNA sequence, 5\u2019-GGTCTC-3\u2019, and cleaves it away from the recognition site at positions 1\/5: 5\u2019-GGTCTC(N)1^\/3\u2019-CCAGAG(N)5^-5\u2019. Thus, Pse31I is a true isoschizomer of the restriction enzyme Eco31I [1].
\nThe producer strain was identified based on morphological and biochemical characteristics, as well as by analyzing the primary structure of a fragment of the 16S rRNA gene. Pse31I exhibits maximal activity at 37\u00b0C and is inactivated by incubation at 55\u00b0C and 65\u00b0C. The enzyme does not cleave sites containing a methylated cytosine in the top strand (5\u2019-GGTCT(5mC)-3\u2019) and\/or in the bottom strand (3\u2019-(5mC)CAGAG).
\nThe optimal reaction conditions for the enzyme include SE-buffer 5 (Y) (33 mM Tris-acetate (pH 7.9), 10 mM Mg-acetate, 66 mM K-acetate, 1 mM DTT) and a temperature of 37\u00b0C<\/p>\n","protected":false},"author":1,"featured_media":427,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[5],"tags":[44],"coauthors":[8,12,13,43,10,9,16],"class_list":["post-400","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-new-enzymes","tag-pse31i"],"_links":{"self":[{"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/posts\/400","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/comments?post=400"}],"version-history":[{"count":16,"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/posts\/400\/revisions"}],"predecessor-version":[{"id":423,"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/posts\/400\/revisions\/423"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/media\/427"}],"wp:attachment":[{"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/media?parent=400"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/categories?post=400"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/tags?post=400"},{"taxonomy":"author","embeddable":true,"href":"https:\/\/dnape.online\/en\/wp-json\/wp\/v2\/coauthors?post=400"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}