{"id":"https://openalex.org/W3144845331","doi":"https://doi.org/10.21203/rs.3.rs-91180/v1","title":"High Cell Density Cultivation of A Recombinant Bacillus Subtilis for Nattokinase Production","display_name":"High Cell Density Cultivation of A Recombinant Bacillus Subtilis for Nattokinase Production","publication_year":2020,"publication_date":"2020-10-16","ids":{"openalex":"https://openalex.org/W3144845331","doi":"https://doi.org/10.21203/rs.3.rs-91180/v1","mag":"3144845331"},"language":"en","primary_location":{"id":"doi:10.21203/rs.3.rs-91180/v1","doi":"https://doi.org/10.21203/rs.3.rs-91180/v1","is_oa":true,"landing_page_url":"https://doi.org/10.21203/rs.3.rs-91180/v1","pdf_url":"https://www.researchsquare.com/article/rs-91180/latest.pdf","source":null,"license":"cc-by","license_id":"https://openalex.org/licenses/cc-by","version":"submittedVersion","is_accepted":false,"is_published":false,"raw_source_name":null,"raw_type":"posted-content"},"volume":null,"type":"preprint","is_traditional":true,"indexed_in":["crossref"],"open_access":{"is_oa":true,"oa_status":"green","oa_url":"https://www.researchsquare.com/article/rs-91180/latest.pdf","any_repository_has_fulltext":null},"authorships":[{"author_position":"first","author":{"id":"https://openalex.org/A5100434772","display_name":"Jianhua Zhang","orcid":"https://orcid.org/0000-0002-8097-4790","observed_orcids":["https://orcid.org/0000-0002-8097-4790"]},"institutions":[{"id":"https://openalex.org/I183067930","display_name":"Shanghai Jiao Tong University","ror":"https://ror.org/0220qvk04","country_code":"CN","type":"education","lineage":["https://openalex.org/I183067930"]}],"countries":["CN"],"is_corresponding":true,"raw_author_name":"Jianhua Zhang","raw_affiliation_strings":["Shanghai Jiao Tong University","Shanghai Jiao Tong University Research","University"],"raw_orcid":"https://orcid.org/0000-0002-8097-4790","affiliations":[{"raw_affiliation_string":"Shanghai Jiao Tong University","institution_ids":["https://openalex.org/I183067930"]},{"raw_affiliation_string":"Shanghai Jiao Tong University Research","institution_ids":["https://openalex.org/I183067930"]},{"raw_affiliation_string":"University","institution_ids":[]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5100916236","display_name":"Qing Cui","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I183067930","display_name":"Shanghai Jiao Tong University","ror":"https://ror.org/0220qvk04","country_code":"CN","type":"education","lineage":["https://openalex.org/I183067930"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Qing Cui","raw_affiliation_strings":["Shanghai Jiao Tong University","Shanghai Jiao Tong University Research","University"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Shanghai Jiao Tong University","institution_ids":["https://openalex.org/I183067930"]},{"raw_affiliation_string":"Shanghai Jiao Tong University Research","institution_ids":["https://openalex.org/I183067930"]},{"raw_affiliation_string":"University","institution_ids":[]}]},{"author_position":"middle","author":{"id":"https://openalex.org/A5053137351","display_name":"Bingjun Qian","orcid":"https://orcid.org/0000-0003-2490-9596","observed_orcids":["https://orcid.org/0000-0003-2490-9596"]},"institutions":[{"id":"https://openalex.org/I183067930","display_name":"Shanghai Jiao Tong University","ror":"https://ror.org/0220qvk04","country_code":"CN","type":"education","lineage":["https://openalex.org/I183067930"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Bingjun Qian","raw_affiliation_strings":["Shanghai Jiao Tong University","Shanghai Jiao Tong University Research","University"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Shanghai Jiao Tong University","institution_ids":["https://openalex.org/I183067930"]},{"raw_affiliation_string":"Shanghai Jiao Tong University Research","institution_ids":["https://openalex.org/I183067930"]},{"raw_affiliation_string":"University","institution_ids":[]}]},{"author_position":"last","author":{"id":"https://openalex.org/A5100801760","display_name":"Xiangjun Sun","orcid":null,"observed_orcids":[]},"institutions":[{"id":"https://openalex.org/I183067930","display_name":"Shanghai Jiao Tong University","ror":"https://ror.org/0220qvk04","country_code":"CN","type":"education","lineage":["https://openalex.org/I183067930"]}],"countries":["CN"],"is_corresponding":false,"raw_author_name":"Xiangjun Sun","raw_affiliation_strings":["Shanghai Jiao Tong University","Shanghai Jiao Tong University Research","University"],"raw_orcid":null,"affiliations":[{"raw_affiliation_string":"Shanghai Jiao Tong University","institution_ids":["https://openalex.org/I183067930"]},{"raw_affiliation_string":"Shanghai Jiao Tong University Research","institution_ids":["https://openalex.org/I183067930"]},{"raw_affiliation_string":"University","institution_ids":[]}]}],"institutions":[],"countries_distinct_count":1,"institutions_distinct_count":1,"corresponding_author_ids":["https://openalex.org/A5100434772"],"corresponding_institution_ids":["https://openalex.org/I183067930"],"apc_list":null,"apc_paid":null,"fwci":null,"has_fulltext":true,"cited_by_count":3,"citation_normalized_percentile":null,"cited_by_percentile_year":null,"biblio":{"volume":null,"issue":null,"first_page":null,"last_page":null},"is_retracted":false,"is_paratext":false,"is_xpac":false,"primary_topic":{"id":"https://openalex.org/T11617","display_name":"Enzyme Production and Characterization","score":0.9965999722480774,"subfield":{"id":"https://openalex.org/subfields/1305","display_name":"Biotechnology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},"topics":[{"id":"https://openalex.org/T11617","display_name":"Enzyme Production and Characterization","score":0.9965999722480774,"subfield":{"id":"https://openalex.org/subfields/1305","display_name":"Biotechnology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},{"id":"https://openalex.org/T10497","display_name":"Fungal and yeast genetics research","score":0.0007999999797903001,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},{"id":"https://openalex.org/T10932","display_name":"Microbial Metabolic Engineering and Bioproduction","score":0.0005000000237487257,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}}],"keywords":[{"id":"https://openalex.org/keywords/nattokinase","display_name":"nattokinase","score":1.0},{"id":"https://openalex.org/keywords/recombinant-bacillus-subtilis","display_name":"recombinant Bacillus subtilis","score":0.9769999980926514},{"id":"https://openalex.org/keywords/high-cell-density-culture","display_name":"high cell density culture","score":0.8289999961853027},{"id":"https://openalex.org/keywords/fed-batch-fermentation","display_name":"fed-batch fermentation","score":0.996999979019165},{"id":"https://openalex.org/keywords/induction-timing","display_name":"induction timing","score":0.515999972820282},{"id":"https://openalex.org/keywords/feeding-strategies","display_name":"feeding strategies","score":0.7680000066757202},{"id":"https://openalex.org/keywords/fibrinolytic-enzyme","display_name":"fibrinolytic enzyme","score":0.7689999938011169},{"id":"https://openalex.org/keywords/enzyme-purification","display_name":"enzyme purification","score":0.7200000286102295},{"id":"https://openalex.org/keywords/ammonium-sulfate-precipitation","display_name":"ammonium sulfate precipitation","score":0.8220000267028809},{"id":"https://openalex.org/keywords/nickel-affinity-chromatography","display_name":"nickel affinity chromatography","score":0.9710000157356262},{"id":"https://openalex.org/keywords/nutraceuticals","display_name":"nutraceuticals","score":0.7710000276565552}],"concepts":[{"id":"https://openalex.org/C2778568395","wikidata":"https://www.wikidata.org/wiki/Q11323875","display_name":"Nattokinase","level":3,"score":0.9680611491203308},{"id":"https://openalex.org/C2777272437","wikidata":"https://www.wikidata.org/wiki/Q131238","display_name":"Bacillus subtilis","level":3,"score":0.9074183702468872},{"id":"https://openalex.org/C100544194","wikidata":"https://www.wikidata.org/wiki/Q41760","display_name":"Fermentation","level":2,"score":0.620160698890686},{"id":"https://openalex.org/C2776428949","wikidata":"https://www.wikidata.org/wiki/Q18814659","display_name":"Ammonium sulfate precipitation","level":4,"score":0.6192664504051208},{"id":"https://openalex.org/C40767141","wikidata":"https://www.wikidata.org/wiki/Q285697","display_name":"Recombinant DNA","level":3,"score":0.5851002335548401},{"id":"https://openalex.org/C198684833","wikidata":"https://www.wikidata.org/wiki/Q2823748","display_name":"Specific activity","level":3,"score":0.5387709140777588},{"id":"https://openalex.org/C28212737","wikidata":"https://www.wikidata.org/wiki/Q5381543","display_name":"Enzyme assay","level":3,"score":0.5235871076583862},{"id":"https://openalex.org/C181199279","wikidata":"https://www.wikidata.org/wiki/Q8047","display_name":"Enzyme","level":2,"score":0.5128381848335266},{"id":"https://openalex.org/C185592680","wikidata":"https://www.wikidata.org/wiki/Q2329","display_name":"Chemistry","level":0,"score":0.511011004447937},{"id":"https://openalex.org/C57409179","wikidata":"https://www.wikidata.org/wiki/Q1218293","display_name":"Lysis","level":2,"score":0.46542876958847046},{"id":"https://openalex.org/C43617362","wikidata":"https://www.wikidata.org/wiki/Q170050","display_name":"Chromatography","level":1,"score":0.46493786573410034},{"id":"https://openalex.org/C55493867","wikidata":"https://www.wikidata.org/wiki/Q7094","display_name":"Biochemistry","level":1,"score":0.44942331314086914},{"id":"https://openalex.org/C2780925461","wikidata":"https://www.wikidata.org/wiki/Q191831","display_name":"Ammonium sulfate","level":2,"score":0.43093764781951904},{"id":"https://openalex.org/C31903555","wikidata":"https://www.wikidata.org/wiki/Q1637030","display_name":"Food science","level":1,"score":0.38774195313453674},{"id":"https://openalex.org/C86803240","wikidata":"https://www.wikidata.org/wiki/Q420","display_name":"Biology","level":0,"score":0.3754953444004059},{"id":"https://openalex.org/C523546767","wikidata":"https://www.wikidata.org/wiki/Q10876","display_name":"Bacteria","level":2,"score":0.20345163345336914},{"id":"https://openalex.org/C104317684","wikidata":"https://www.wikidata.org/wiki/Q7187","display_name":"Gene","level":2,"score":0.14665848016738892},{"id":"https://openalex.org/C54355233","wikidata":"https://www.wikidata.org/wiki/Q7162","display_name":"Genetics","level":1,"score":0.0},{"id":"https://openalex.org/C193363816","wikidata":"https://www.wikidata.org/wiki/Q1617865","display_name":"Size-exclusion chromatography","level":3,"score":0.0}],"mesh":[],"locations_count":1,"locations":[{"id":"doi:10.21203/rs.3.rs-91180/v1","doi":"https://doi.org/10.21203/rs.3.rs-91180/v1","is_oa":true,"landing_page_url":"https://doi.org/10.21203/rs.3.rs-91180/v1","pdf_url":"https://www.researchsquare.com/article/rs-91180/latest.pdf","source":null,"license":"cc-by","license_id":"https://openalex.org/licenses/cc-by","version":"submittedVersion","is_accepted":false,"is_published":false,"raw_source_name":null,"raw_type":"posted-content"}],"best_oa_location":{"id":"doi:10.21203/rs.3.rs-91180/v1","doi":"https://doi.org/10.21203/rs.3.rs-91180/v1","is_oa":true,"landing_page_url":"https://doi.org/10.21203/rs.3.rs-91180/v1","pdf_url":"https://www.researchsquare.com/article/rs-91180/latest.pdf","source":null,"license":"cc-by","license_id":"https://openalex.org/licenses/cc-by","version":"submittedVersion","is_accepted":false,"is_published":false,"raw_source_name":null,"raw_type":"posted-content"},"sustainable_development_goals":[{"display_name":"Zero hunger","score":0.40450000762939453,"id":"https://metadata.un.org/sdg/2"}],"study_designs":[],"awards":[{"id":"https://openalex.org/G6030949499","display_name":null,"funder_award_id":"2017YFD0400205","funder_id":"https://openalex.org/F4320335777","funder_display_name":"National Key Research and Development Program of China"}],"funders":[{"id":"https://openalex.org/F4320335777","display_name":"National Key Research and Development Program of China","ror":null}],"has_content":{"pdf":true,"grobid_xml":true},"content_urls":{"pdf":"https://content.openalex.org/works/W3144845331.pdf","grobid_xml":"https://content.openalex.org/works/W3144845331.grobid-xml"},"referenced_works_count":23,"referenced_works":["https://openalex.org/W1701939279","https://openalex.org/W1942265794","https://openalex.org/W1976577891","https://openalex.org/W1976802074","https://openalex.org/W1983501453","https://openalex.org/W1994370219","https://openalex.org/W2005127242","https://openalex.org/W2006080326","https://openalex.org/W2014127565","https://openalex.org/W2052886960","https://openalex.org/W2055209600","https://openalex.org/W2057848601","https://openalex.org/W2075341454","https://openalex.org/W2081375075","https://openalex.org/W2081673601","https://openalex.org/W2088051109","https://openalex.org/W2101108802","https://openalex.org/W2141622176","https://openalex.org/W2406012908","https://openalex.org/W2786774014","https://openalex.org/W2894600673","https://openalex.org/W2944014215","https://openalex.org/W2947240423"],"related_works":["https://openalex.org/W4293063911","https://openalex.org/W2184913856","https://openalex.org/W2006080326","https://openalex.org/W2055209600","https://openalex.org/W2947240423","https://openalex.org/W2385310165","https://openalex.org/W4409986443","https://openalex.org/W2252721507","https://openalex.org/W4397036677","https://openalex.org/W1976577891"],"abstract_inverted_index":{"Abstract":[0],"Background:":[1],"Nattokinase":[2],"(NK),":[3],"a":[4,19,25,28,32,42,47,52,67,83,97,114,145,188,201,218,236],"fibrinolytic":[5],"enzyme,":[6],"can":[7,224,243],"be":[8,225,244],"produced":[9],"by":[10,165,178,227],"culturing":[11],"recombinant":[12,212],"Bacillus":[13,213],"subtilis":[14,49,214],"in":[15,18],"Luria-Bertani":[16],"broth":[17],"shaking":[20],"flask.":[21],"For":[22,61,134],"use":[23],"as":[24],"nutraceutical,":[26],"however,":[27,125],"large-scale":[29],"preparation":[30],"and":[31,82,100,109,143,182,208,231,234,240],"simple":[33],"purification":[34],"process":[35,44],"are":[36],"required.Results:":[37],"The":[38,221],"present":[39],"study":[40],"utilized":[41],"fed-batch":[43,219],"to":[45,104,156],"cultivate":[46],"B.":[48],"strain":[50],"carrying":[51],"pHT01":[53],"plasmid":[54],"with":[55,66,96,187,217],"an":[56,74],"NK-encoding":[57],"gene":[58],"(B.":[59],"subtilis/pHT01-aprN1).":[60],"batch":[62],"A":[63],"(FB":[64],"A),":[65],"pH-stat":[68],"two-stage":[69],"fermentation":[70,173],"strategy,":[71,233],"we":[72,92],"achieved":[73],"activity":[75,85,116,128,147,163,242],"of":[76,86,117,148,193],"2910.5":[77],"\u00b1":[78,119],"21.6":[79],"U":[80,88,121,151],"mL-1":[81,122,152],"specific":[84,127],"30.32":[87],"ml-1":[89],"OD600-1.":[90],"Then,":[91],"changed":[93],"the":[94,126,135,161,166,204,229],"strategy":[95,140],"later":[98],"induction":[99,230],"lower":[101,130],"feeding":[102,139,232],"rate":[103,192,207],"pursue":[105],"higher":[106,111],"cell":[107,205,238],"density":[108,239],"thus":[110,235],"enzyme":[112,241],"activity,":[113],"11.9-fold":[115],"4521.8":[118],"23.8":[120],"was":[123,129,141,153,160,175],"acquired,":[124],"than":[131],"FB":[132],"A.":[133],"third":[136],"batch,":[137],"low-glycerol-level-maintain":[138],"followed,":[142],"finally,":[144],"NK":[146,190,209],"7778":[149],"\u00b117.28":[150],"obtained,":[154],"according":[155],"our":[157],"knowledge,":[158],"it":[159],"highest":[162],"assayed":[164],"fibrin":[167],"plate":[168],"method":[169],"ever":[170],"reported.":[171],"Furthermore,":[172],"supernatant":[174],"successively":[176],"purified":[177],"ammonium":[179],"sulfate":[180],"precipitation":[181],"nickel":[183],"column":[184],"affinity":[185],"chromatography":[186],"total":[189],"recovery":[191],"65.2%.Conclusions:":[194],"Our":[195],"results":[196],"indicate":[197],"that":[198],"there":[199],"is":[200,215],"balance":[202],"between":[203],"growth":[206],"expression":[210],"when":[211],"cultured":[216],"process.":[220],"equilibrium":[222],"state":[223],"attained":[226],"optimizing":[228],"high":[237],"achieved.":[245]},"counts_by_year":[{"year":2026,"cited_by_count":1},{"year":2025,"cited_by_count":1},{"year":2022,"cited_by_count":1}],"updated_date":"2026-11-09T13:22:03.289127","created_date":"2021-04-13T00:00:00"}
