K Koga, Y Iwamoto, H Sakamoto, K Hatano, M Sano, I Kato
beta-N-Acetylhexosaminidase was produced by Trichoderma harzianum cultivated with chitin as the growth substrate. The enzyme was purified 13.2-fold to homogeneity by ultrafiltration and sequential chromatography on SP-Toyopearl and Sephacryl S-200. The molecular weight of the enzyme was estimated to be about 150,000 by gel filtration. The pH and temperature optima were 4.0-5.5 and 50 degrees C, respectively. The enzyme hydrolyzed N-acetylchitooligosaccharides at the non-reducing ends to release GlcNAc monomer. The enzyme showed a strict substrate specificity to the sugar chains in complex carbohydrates, hydrolyzing only the linkage of GlcNAc beta 1-3Gal, but not hydrolyzing the other linkages such as GalNAc beta 1-3Gal and GlcNAc beta 1-2Man.
{"title":"Purification and characterization of beta-N-acetylhexosaminidase from Trichoderma harzianum.","authors":"K Koga, Y Iwamoto, H Sakamoto, K Hatano, M Sano, I Kato","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>beta-N-Acetylhexosaminidase was produced by Trichoderma harzianum cultivated with chitin as the growth substrate. The enzyme was purified 13.2-fold to homogeneity by ultrafiltration and sequential chromatography on SP-Toyopearl and Sephacryl S-200. The molecular weight of the enzyme was estimated to be about 150,000 by gel filtration. The pH and temperature optima were 4.0-5.5 and 50 degrees C, respectively. The enzyme hydrolyzed N-acetylchitooligosaccharides at the non-reducing ends to release GlcNAc monomer. The enzyme showed a strict substrate specificity to the sugar chains in complex carbohydrates, hydrolyzing only the linkage of GlcNAc beta 1-3Gal, but not hydrolyzing the other linkages such as GalNAc beta 1-3Gal and GlcNAc beta 1-2Man.</p>","PeriodicalId":7729,"journal":{"name":"Agricultural and biological chemistry","volume":"55 11","pages":"2817-23"},"PeriodicalIF":0.0,"publicationDate":"1991-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12540514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hiroshi Shinmoto, S. Dosako, Hirofumi Tachibana, S. Shirahata, H. Murakami
antibodies are required for the therapy of diseases to avoid undesirable side effects.9' Recently, we developed human-mouse hybridomas secreting human IgM class anti-ricin or anti-diphtheria toxin antibodies by the transformation of human peripheral blood lymphocytes (PBLs) with Epstein-Barr virus (EBV), followed by cell fusion between the transformed cells and mousemyelomaSP2/O2.10) With these hybridomas, we established hybrid hybridomas that secreted IgM class human bifunctional antibodies as a model experiment.ll' However, the affinity of the secreted hybrid IgM to diphtheria toxin was lower than that to ricin. One possibility for this is the low expression of immunoglobulin polypeptide genes to diphtheria toxin in the hybrid hybridoma, due probably to improper technique for the generation of the hybrid hybridoma. The previous method10' used actinomycin-D treatment of one hybridoma line to fuse with another 6-thioguanineresistant hybridoma line. Wesupposed that the actinomycin-D treatment in the fusion procedure were responsible for the lower affinity to diphtheria toxin of the produced bifunctional antibody, because Kobayashi et al. 12) reported that a T-cell hybridoma that had been generated by the actinomycin-D method tended to lose its capability of secreting some lymphokines. To solve this problem, we Fig. 1. Scheme for Generation of Hybrid Hybridoma Secreting Anti-neocarzinostatin/Anti-diphtheria Toxin Bifunctional Antibody.
{"title":"Generation of human-mouse hybridoma secreting human IgM class anti-neocarzinostatin antibody and its application to hybrid hybridoma.","authors":"Hiroshi Shinmoto, S. Dosako, Hirofumi Tachibana, S. Shirahata, H. Murakami","doi":"10.1271/BBB1961.55.2883","DOIUrl":"https://doi.org/10.1271/BBB1961.55.2883","url":null,"abstract":"antibodies are required for the therapy of diseases to avoid undesirable side effects.9' Recently, we developed human-mouse hybridomas secreting human IgM class anti-ricin or anti-diphtheria toxin antibodies by the transformation of human peripheral blood lymphocytes (PBLs) with Epstein-Barr virus (EBV), followed by cell fusion between the transformed cells and mousemyelomaSP2/O2.10) With these hybridomas, we established hybrid hybridomas that secreted IgM class human bifunctional antibodies as a model experiment.ll' However, the affinity of the secreted hybrid IgM to diphtheria toxin was lower than that to ricin. One possibility for this is the low expression of immunoglobulin polypeptide genes to diphtheria toxin in the hybrid hybridoma, due probably to improper technique for the generation of the hybrid hybridoma. The previous method10' used actinomycin-D treatment of one hybridoma line to fuse with another 6-thioguanineresistant hybridoma line. Wesupposed that the actinomycin-D treatment in the fusion procedure were responsible for the lower affinity to diphtheria toxin of the produced bifunctional antibody, because Kobayashi et al. 12) reported that a T-cell hybridoma that had been generated by the actinomycin-D method tended to lose its capability of secreting some lymphokines. To solve this problem, we Fig. 1. Scheme for Generation of Hybrid Hybridoma Secreting Anti-neocarzinostatin/Anti-diphtheria Toxin Bifunctional Antibody.","PeriodicalId":7729,"journal":{"name":"Agricultural and biological chemistry","volume":"133 1","pages":"2883-5"},"PeriodicalIF":0.0,"publicationDate":"1991-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79374760","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1991-11-01DOI: 10.1080/00021369.1991.10871053
S. Iijima, Mitsuyoshi Ishida, S. Nakajima‐Iijima, T. Hishida, Hideki Watanabe, Takeshi Kobayashi
Human endothelial cells isolated from an umbilical cord vein were transfected with origin-defective simian virus 40 (SV40) DNA. Among several of the SV40 transfected clones isolated, cell lines SV-2 and SV-3 showed a normal endothelial cell morphology and extended life span, and could survive almost 100 generations. Just before crisis, the morphology of SV-3 changed. SV-3T cell line was isolated from this SV-3 culture, which acquired an almost infinite life span, rapid growth rate and the ability to grow in soft agar. At the same time, the SV-3T cell line lost the factor VIII-related antigen and normal endothelial cell morphology, and showed an abnormal chromosome number. Further characterization showed the ability of SV-2 and SV-3T to produce increasing amounts of tissue plasminogen activator and a similar level of a plasminogen activator inhibitor compared with normal human endothelial cells. These results indicate that the SV-3T cell line was transformed and acquired an infinite life span while still r...
{"title":"Immortalization of human endothelial cells by origin-defective simian virus 40 DNA","authors":"S. Iijima, Mitsuyoshi Ishida, S. Nakajima‐Iijima, T. Hishida, Hideki Watanabe, Takeshi Kobayashi","doi":"10.1080/00021369.1991.10871053","DOIUrl":"https://doi.org/10.1080/00021369.1991.10871053","url":null,"abstract":"Human endothelial cells isolated from an umbilical cord vein were transfected with origin-defective simian virus 40 (SV40) DNA. Among several of the SV40 transfected clones isolated, cell lines SV-2 and SV-3 showed a normal endothelial cell morphology and extended life span, and could survive almost 100 generations. Just before crisis, the morphology of SV-3 changed. SV-3T cell line was isolated from this SV-3 culture, which acquired an almost infinite life span, rapid growth rate and the ability to grow in soft agar. At the same time, the SV-3T cell line lost the factor VIII-related antigen and normal endothelial cell morphology, and showed an abnormal chromosome number. Further characterization showed the ability of SV-2 and SV-3T to produce increasing amounts of tissue plasminogen activator and a similar level of a plasminogen activator inhibitor compared with normal human endothelial cells. These results indicate that the SV-3T cell line was transformed and acquired an infinite life span while still r...","PeriodicalId":7729,"journal":{"name":"Agricultural and biological chemistry","volume":"6 1","pages":"2847-2853"},"PeriodicalIF":0.0,"publicationDate":"1991-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82090856","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1991-11-01DOI: 10.1080/00021369.1991.10871046
Akiyoshi Tanaka
{"title":"Differential Scanning Calorimetrie Study of the Thermal Denaturation of Almondβ-Glucosidase","authors":"Akiyoshi Tanaka","doi":"10.1080/00021369.1991.10871046","DOIUrl":"https://doi.org/10.1080/00021369.1991.10871046","url":null,"abstract":"","PeriodicalId":7729,"journal":{"name":"Agricultural and biological chemistry","volume":"14 1","pages":"2773-2776"},"PeriodicalIF":0.0,"publicationDate":"1991-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89090051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yukiharu Sato, T. Kojima, T. Goto, R. Oomikawa, Hiroyuki Watanabe, K. Wakabayashi
In order to explain the close phytotoxic activities of N-aryl-3,4,5,6-tetrahydrophthalimides and their hydrolyzed products, N-aryl-3,4,5,6-tetrahydrophthalamic acids, five sets of both types of compounds possessing the same aryl residues were prepared. Their phytotoxic activity against sawa millett (E. utitts) and green microalga (S. acutus), and the relationship of interconversion between the imides and amide acids during the bioassay were investigated. In almost all cases, the imides showed stronger activity than the corresponding amide acids. The hydrolysis of the imides and the cyclization of amide acids were observed in respect of the aryl residues. The phytotoxicity caused by the imides and amide acids tested was influenced by this interconversion.
{"title":"Hydrolysis and Phytotoxic Activity of Cyclic Imides","authors":"Yukiharu Sato, T. Kojima, T. Goto, R. Oomikawa, Hiroyuki Watanabe, K. Wakabayashi","doi":"10.1271/BBB1961.55.2677","DOIUrl":"https://doi.org/10.1271/BBB1961.55.2677","url":null,"abstract":"In order to explain the close phytotoxic activities of N-aryl-3,4,5,6-tetrahydrophthalimides and their hydrolyzed products, N-aryl-3,4,5,6-tetrahydrophthalamic acids, five sets of both types of compounds possessing the same aryl residues were prepared. Their phytotoxic activity against sawa millett (E. utitts) and green microalga (S. acutus), and the relationship of interconversion between the imides and amide acids during the bioassay were investigated. In almost all cases, the imides showed stronger activity than the corresponding amide acids. The hydrolysis of the imides and the cyclization of amide acids were observed in respect of the aryl residues. The phytotoxicity caused by the imides and amide acids tested was influenced by this interconversion.","PeriodicalId":7729,"journal":{"name":"Agricultural and biological chemistry","volume":"33 1","pages":"2677-2681"},"PeriodicalIF":0.0,"publicationDate":"1991-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80059917","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The application of both pressurization and enzymatic treatment to aged rice grains was investigated to improve their cooking properties. Pressurization at 100 MPa was favorable in improving the properties. For treatment of the grains with enzymes, actinase was more effective than cellulase and pectolyase. Neither lipase nor transglutaminase showed any imporving effect. Actinase-treated grains, when cooked, gave the most farvorable result in terms of the stickiness/hardness ratio, brightness, flavor, and texture.
{"title":"Improving the cooking properties of aged rice grains by pressurization and enzymatic treatment","authors":"Michiko Watanabe, Eiko Arai, Kazuo Honma, S. Fuke","doi":"10.1271/BBB1961.55.2725","DOIUrl":"https://doi.org/10.1271/BBB1961.55.2725","url":null,"abstract":"The application of both pressurization and enzymatic treatment to aged rice grains was investigated to improve their cooking properties. Pressurization at 100 MPa was favorable in improving the properties. For treatment of the grains with enzymes, actinase was more effective than cellulase and pectolyase. Neither lipase nor transglutaminase showed any imporving effect. Actinase-treated grains, when cooked, gave the most farvorable result in terms of the stickiness/hardness ratio, brightness, flavor, and texture.","PeriodicalId":7729,"journal":{"name":"Agricultural and biological chemistry","volume":"94 1 1","pages":"2725-2731"},"PeriodicalIF":0.0,"publicationDate":"1991-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87666710","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The Pseudomonas fluorescens gene (estB) that encodes a novel esterase (esterase II) was cloned into Escherichia coli JM83. DNA sequencing found a single open reading frame of 654 nucleotides. The open reading frame was confirmed by N-terminal amino acid sequence analysis of the esterase protein. A potential Shine-Dalgarno sequence is followed by the coding sequence of the estB gene. The amino acid sequence deduced from the nucleotide sequence contains the consensus active site sequence, G-X-S-X-G, of serine esterases. The enzyme expressed in an E. coli clone was purified by ion-exchange chromatography and gel filtration. Homogeneity of the purified enzyme was confirmed using SDS-polyacrylamide gel electrophoresis. The native enzyme exists as a dimer consisting of two identical subunits, each with a molecular weight of 23,000. The results of the experiments for identifying substrate specificity and the inhibitor studies suggest that this enzyme is a carboxylesterase (EC 3.1.1.1) and a serine residue is present at the active site of the esterase, as in the esterases of animal tissues.
{"title":"Characterization of Pseudomonas fluorescens carboxylesterase: cloning and expression of the esterase gene in Escherichia coli.","authors":"K H Hong, W H Jang, K D Choi, O J Yoo","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The Pseudomonas fluorescens gene (estB) that encodes a novel esterase (esterase II) was cloned into Escherichia coli JM83. DNA sequencing found a single open reading frame of 654 nucleotides. The open reading frame was confirmed by N-terminal amino acid sequence analysis of the esterase protein. A potential Shine-Dalgarno sequence is followed by the coding sequence of the estB gene. The amino acid sequence deduced from the nucleotide sequence contains the consensus active site sequence, G-X-S-X-G, of serine esterases. The enzyme expressed in an E. coli clone was purified by ion-exchange chromatography and gel filtration. Homogeneity of the purified enzyme was confirmed using SDS-polyacrylamide gel electrophoresis. The native enzyme exists as a dimer consisting of two identical subunits, each with a molecular weight of 23,000. The results of the experiments for identifying substrate specificity and the inhibitor studies suggest that this enzyme is a carboxylesterase (EC 3.1.1.1) and a serine residue is present at the active site of the esterase, as in the esterases of animal tissues.</p>","PeriodicalId":7729,"journal":{"name":"Agricultural and biological chemistry","volume":"55 11","pages":"2839-45"},"PeriodicalIF":0.0,"publicationDate":"1991-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12540515","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H Shinmoto, S Dosako, H Tachibana, S Shirahata, H Murakami
{"title":"Generation of human-mouse hybridoma secreting human IgM class anti-neocarzinostatin antibody and its application to hybrid hybridoma.","authors":"H Shinmoto, S Dosako, H Tachibana, S Shirahata, H Murakami","doi":"","DOIUrl":"","url":null,"abstract":"","PeriodicalId":7729,"journal":{"name":"Agricultural and biological chemistry","volume":"55 11","pages":"2883-5"},"PeriodicalIF":0.0,"publicationDate":"1991-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12540517","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Treatment of grayanotoxin (G) III with pyruvic acid in methanol gave the Δ1(10)-1,5-seco-G derivative. Photo-sensitized oxygenation of Δ1 (10)-1 ,5-seco-G under UV light irradiation in methanol gave the 1(S)-perhydroxy- and 1(S)-hydroxy-1,5-seco-G derivatives. The IR and NMR data for Δ1(10)-1(S)-hydroxy-1,5-seco-G were identical with those of natural grayanol B described in the literature.
{"title":"Transformation of Grayanotoxin III to the 1, 5-seco-Grayanotoxin Derivative, Grayanol B","authors":"T. Terai, K. Kiyono, K. Gotō","doi":"10.1271/BBB1961.55.2711","DOIUrl":"https://doi.org/10.1271/BBB1961.55.2711","url":null,"abstract":"Treatment of grayanotoxin (G) III with pyruvic acid in methanol gave the Δ1(10)-1,5-seco-G derivative. Photo-sensitized oxygenation of Δ1 (10)-1 ,5-seco-G under UV light irradiation in methanol gave the 1(S)-perhydroxy- and 1(S)-hydroxy-1,5-seco-G derivatives. The IR and NMR data for Δ1(10)-1(S)-hydroxy-1,5-seco-G were identical with those of natural grayanol B described in the literature.","PeriodicalId":7729,"journal":{"name":"Agricultural and biological chemistry","volume":"77 1","pages":"2711-2715"},"PeriodicalIF":0.0,"publicationDate":"1991-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87058242","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Essential Role of Phosphorylation in Signal Transduction by a Chimeric Receptor Composed of the Chemoreceptor Tar and Osmosensor EnvZ","authors":"R. Utsumi, S. Forst, M. Noda","doi":"10.1271/BBB1961.55.2897","DOIUrl":"https://doi.org/10.1271/BBB1961.55.2897","url":null,"abstract":"","PeriodicalId":7729,"journal":{"name":"Agricultural and biological chemistry","volume":"110 1","pages":"2897-2898"},"PeriodicalIF":0.0,"publicationDate":"1991-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84379682","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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