{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T00:56:22Z","timestamp":1777596982683,"version":"3.51.4"},"reference-count":65,"publisher":"American Society for Microbiology","issue":"12","license":[{"start":{"date-parts":[[2011,6,15]],"date-time":"2011-06-15T00:00:00Z","timestamp":1308096000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2011,6,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Trehalose accumulation is a common cell defense strategy against a variety of stressful conditions. In particular, our team detected high levels of trehalose in<jats:named-content content-type=\"genus-species\">Propionibacterium freudenreichii<\/jats:named-content>in response to acid stress, a result that led to the idea that endowing<jats:named-content content-type=\"genus-species\">Lactococcus lactis<\/jats:named-content>with the capacity to synthesize trehalose could improve the acid tolerance of this organism. To this end, we took advantage of the endogenous genes involved in the trehalose catabolic pathway of<jats:named-content content-type=\"genus-species\">L. lactis<\/jats:named-content>, i.e.,<jats:italic>trePP<\/jats:italic>and<jats:italic>pgmB<\/jats:italic>, encoding trehalose 6-phosphate phosphorylase and \u03b2-phosphoglucomutase, respectively, which enabled the synthesis of trehalose 6-phosphate. Given that<jats:named-content content-type=\"genus-species\">L. lactis<\/jats:named-content>lacks trehalose 6-phosphate phosphatase, the respective gene,<jats:italic>otsB<\/jats:italic>, from the food-grade organism<jats:named-content content-type=\"genus-species\">P. freudenreichii<\/jats:named-content>was used to provide the required activity. The trehalose yield was approximately 15% in resting cells and in mid-exponential-phase cells grown without pH control. The intracellular concentration of trehalose reached maximal values of approximately 170 mM, but at least 67% of the trehalose produced was found in the growth medium. The viability of mutant and control strains was examined after exposure to heat, cold or acid shock, and freeze-drying. The trehalose-producing strains showed improved tolerance (5- to 10-fold-higher survivability) to acid (pH 3) and cold shock (4\u00b0C); there was also a strong improvement in cell survival in response to heat shock (45\u00b0C), and no protection was rendered against dehydration. The insight provided by this work may help the design of food-grade strains optimized for the dairy industry as well as for oral drug delivery.<\/jats:p>","DOI":"10.1128\/aem.02922-10","type":"journal-article","created":{"date-parts":[[2011,4,23]],"date-time":"2011-04-23T03:29:25Z","timestamp":1303529365000},"page":"4189-4199","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":54,"title":["Engineering Trehalose Synthesis in Lactococcus lactis for Improved Stress Tolerance"],"prefix":"10.1128","volume":"77","author":[{"given":"Ana L\u00facia","family":"Carvalho","sequence":"first","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica, Universidade Nova de Lisboa, Av. da Rep\u00fablica-EAN, 2780-157 Oeiras, Portugal"}]},{"given":"Filipa S.","family":"Cardoso","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica, Universidade Nova de Lisboa, Av. da Rep\u00fablica-EAN, 2780-157 Oeiras, Portugal"}]},{"given":"Andreas","family":"Bohn","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica, Universidade Nova de Lisboa, Av. da Rep\u00fablica-EAN, 2780-157 Oeiras, Portugal"}]},{"given":"Ana Rute","family":"Neves","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica, Universidade Nova de Lisboa, Av. da Rep\u00fablica-EAN, 2780-157 Oeiras, Portugal"}]},{"given":"Helena","family":"Santos","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica, Universidade Nova de Lisboa, Av. da Rep\u00fablica-EAN, 2780-157 Oeiras, Portugal"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(66)08014-5"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M108279200"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M101487200"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.4161\/hv.5.4.7553"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/7.6.1513"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(76)90527-3"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.200401327"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.29262-0"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1605(03)00387-8"},{"key":"e_1_3_3_11_2","volume-title":"P051. Abstr. 10th Int. Symp. Genet. Ind. Microorgan.","author":"Carvalho A. L.","year":"2006","unstructured":"CarvalhoA. L. . 2006. Towards the production of trehalose in Lactococcus lactis, abstr. P051. Abstr. 10th Int. Symp. Genet. Ind. Microorgan., Prague, Czech Republic."},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1006\/cryo.2000.2260"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.223.4637.701"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.62.10.3662-3667.1996"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibtech.2003.11.011"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19991205)65:5<572::AID-BIT10>3.0.CO;2-7"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/144.3.923"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.84.1.137-146.1962"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M411732200"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1605(03)00104-1"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.252637799"},{"key":"e_1_3_3_22_2","volume-title":"Ph.D. thesis.","author":"Gaspar P","year":"2008","unstructured":"GasparP . 2008. Metabolic engineering of Lactococcus lactis for mannitol production. Ph.D. thesis. ITQB, Universidade Nova de Lisboa, Oeiras, Portugal."},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.154.1.1-9.1983"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1006\/mben.1999.0122"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.184.11.3078-3085.2002"},{"key":"e_1_3_3_26_2","first-page":"195","article-title":"Transformation of Lactococcus by electroporation","volume":"47","author":"Holo H.","year":"1995","unstructured":"HoloH. NesI. F. . 1995. Transformation of Lactococcus by electroporation. Methods Mol. Biol. 47:195\u2013199.","journal-title":"Methods Mol. Biol."},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1474-9726.2010.00582.x"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1023\/A:1020608304886"},{"key":"e_1_3_3_29_2","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1002\/pro.3","article-title":"Effect of trehalose on protein structure","volume":"18","author":"Jain N. K.","year":"2009","unstructured":"JainN. K. RoyI. . 2009. Effect of trehalose on protein structure. Protein Sci. 18:24\u201336.","journal-title":"Protein Sci."},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/0147-619X(92)90022-3"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.142314099"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0958-1669(03)00033-8"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1656(98)00100-X"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1007\/s004380050315"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.61.10.3592-3597.1995"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1605(00)00190-2"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.175.17.5438-5444.1993"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1159\/000106082"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.68.12.6332-6342.2002"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19990720)64:2<200::AID-BIT9>3.0.CO;2-K"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M202573200"},{"key":"e_1_3_3_42_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0723-2020(88)80003-1"},{"key":"e_1_3_3_43_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.1.370-376.2004"},{"key":"e_1_3_3_44_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.7.3845-3854.2004"},{"key":"e_1_3_3_45_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.170.2.700-707.1988"},{"key":"e_1_3_3_46_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.176.17.5304-5311.1994"},{"key":"e_1_3_3_47_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.67.1.33-41.2001"},{"key":"e_1_3_3_48_2","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.26695-0"},{"key":"e_1_3_3_49_2","volume-title":"Molecular cloning: a laboratory manual","author":"Sambrook J.","year":"1989","unstructured":"SambrookJ. FritschE. F. ManiatisT. . 1989. Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.","edition":"2"},{"key":"e_1_3_3_50_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1462-2920.2002.00335.x"},{"key":"e_1_3_3_51_2","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.e04-03-0167"},{"key":"e_1_3_3_52_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7799(98)01251-7"},{"key":"e_1_3_3_53_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.173.3.1187-1192.1991"},{"key":"e_1_3_3_54_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01388-06"},{"key":"e_1_3_3_55_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0968-0004(00)88938-0"},{"key":"e_1_3_3_56_2","doi-asserted-by":"publisher","DOI":"10.1002\/jobm.200900339"},{"key":"e_1_3_3_57_2","doi-asserted-by":"publisher","DOI":"10.1023\/A:1020631532202"},{"key":"e_1_3_3_58_2","doi-asserted-by":"publisher","DOI":"10.1529\/biophysj.106.096461"},{"key":"e_1_3_3_59_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2007.03.011"},{"key":"e_1_3_3_60_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01768-06"},{"key":"e_1_3_3_61_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00395939"},{"key":"e_1_3_3_62_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.57.5.1408-1412.1991"},{"key":"e_1_3_3_63_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0010361"},{"key":"e_1_3_3_64_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.70.11.6738-6747.2004"},{"key":"e_1_3_3_65_2","doi-asserted-by":"publisher","DOI":"10.1242\/jeb.01730"},{"key":"e_1_3_3_66_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01013-06"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.02922-10","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.02922-10","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,4,6]],"date-time":"2024-04-06T09:27:19Z","timestamp":1712395639000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.02922-10"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,6,15]]},"references-count":65,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2011,6,15]]}},"alternative-id":["10.1128\/AEM.02922-10"],"URL":"https:\/\/doi.org\/10.1128\/aem.02922-10","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,6,15]]},"assertion":[{"value":"2010-12-15","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2011-04-09","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2011-06-07","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}