Article
Multidisciplinary Sciences
Eva M. Moya-Gonzalvez, Antonio Rubio-del-Campo, Jesus Rodriguez-Diaz, Maria J. Yebra
Summary: Evidence suggests that HMOs are mainly metabolized by Bifidobacterium species in the infant gut. Bifidobacterium infantis Y538 efficiently consumed all tested HMOs, while other strains only metabolized specific HMOs such as LNT and LNnT.
SCIENTIFIC REPORTS
(2021)
Article
Multidisciplinary Sciences
Zhiqiang Du, Zhengyao Liu, Yinshuang Tan, Kangle Niu, Wei Guo, Yangyang Jia, Xu Fang
Summary: A synergistic strategy utilizing an in vitro multienzyme cascade system was developed to produce LNB, increasing the conversion ratio and yield significantly under optimal conditions. The integration of ATP regeneration and Pi alleviation also decreased the energy required for the reaction by 540 KJ/mol. This approach not only paved the way for producing LNB, but also showed potential for facilitating other chemicals with multienzyme cascades.
Article
Biotechnology & Applied Microbiology
Lijie Chen, Yanmei Qin, Long Ma, Dongdong Meng, Chun You
Summary: A non-ATP-dependent in vitro synthetic enzymatic biosystem was constructed to produce LNB, a core structural unit of HMOs, from starch and GlcNAc. The study demonstrated the feasibility of this biosystem for the synthesis of LNB and shed light on the biosynthesis of other HMOs using low-cost polysaccharides.
SYNTHETIC AND SYSTEMS BIOTECHNOLOGY
(2023)
Article
Agriculture, Multidisciplinary
Fengyi Li, Wei Ma, Lulu Liu, Kangle Niu, Daming Liu, Wencheng Yin, Xudong Zhang, Lijuan Han, Xu Fang
Summary: In this study, LNB was synthesized using a cell factory for the first time. Through the construction of three modules in Kluyveromyces lactis, LNB was successfully produced from lactose and GlcNAc without the addition of cofactors. The engineered K. lactis provides a promising technology platform for the industrial scale production of LNB.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2023)
Article
Food Science & Technology
Xin Huang, Yue Sun, Suhong Li, Motomitsu Kitaoka, Tuoping Li
Summary: To enhance the thermal stability of Lacto-N-biose (LNB), it was encapsulated within OVA-CMC microparticles. With a mass ratio of OVA:CMC:LNB as 1.5:1:1, the microparticles exhibited a particle size of 623 +/- 4 nm, encapsulation efficiency of 83 +/- 1.38%, and loading efficiency of 45 +/- 0.92%. Characterization techniques such as SEM, FT-IR, and XRD confirmed the spherical and uniform distribution of the OVA-CMC-LNB microparticles. Hydrogen bonding and electrostatic interactions were identified as the main forces responsible for the microparticles' assembly. Thermal analysis revealed that the microparticles remained structurally stable at 240 degrees C, greatly improving the thermal stability of LNB. In vitro digestion experiments demonstrated a release rate of LNB from the microparticles of 95 +/- 1.31%. These findings provide a theoretical and technological framework for enhancing the thermal stability of LNB delivery systems.
INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY
(2023)
Article
Food Science & Technology
Wanying Xie, Wenjie Li, Fanhui Gao, Tuoping Li
Summary: Human milk oligosaccharides (HMOs) can reduce early food allergy in infants by improving intestinal flora. Lacto-N-biose (LNB), a component of HMOs, may alleviate infant milk allergy by modulating gut microbiota. In vitro experiments showed that LNB was stable during digestion and fermentation and had a significant impact on the composition of gut microbiota.
INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Wataru Saburi, Tomoya Ota, Koji Kato, Takayoshi Tagami, Keitaro Yamashita, Min Yao, Haruhide Mori
Summary: Lacticaseibacillus casei is a widely used probiotic species that possesses a putative ss-galactosidase. This enzyme hydrolyzes LNB and GNB, but its activity against 6-phospho-ss-galactosidase has not been investigated.
JOURNAL OF APPLIED GLYCOSCIENCE
(2023)
Article
Biochemistry & Molecular Biology
Takayuki Ohnuma, Tomoki Taku, Takeshi Nagatani, Atsushi Horii, Shun Imaoka, Tomonari Tanaka
Summary: The chemo-enzymatic synthesis of lacto-N-biose I (LNB) catalyzed by beta-1,3-galactosidase from Bacillus circulans (BgaC) was achieved using 4,6-dimethoxy-1,3,5-triazin-2-yl beta-galactopyranoside (DMT-beta-Gal) and GlcNAc as substrates. BgaC transferred the Gal moiety to the acceptor to produce LNB, with the maximum yield obtained at a ratio of 1:30 for acceptor to donor substrates.
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
(2021)
Article
Chemistry, Applied
Xiaoxiang Gao, Yinghui Qiu, Luying Gao, Lizhu Zhang, Xiaoqing Li, Yuanyuan Liu, Chao Zhao
Summary: This study demonstrates that LNFPI can inhibit EV71 infection by reducing virus adsorption and promoting the expression of cyclin E, leading to virus-induced S phase arrest and apoptosis inhibition. Moreover, LNFPI can also regulate the abundance of intestinal flora to suppress cell apoptosis. These findings provide new recommendations for the supplementation of LNFPI in infant formula, offering antiviral benefits to formula-fed infants.
Article
Chemistry, Applied
Miaomiao Hu, Mengli Li, Chenchen Li, Tao Zhang
Summary: In this study, an efficient engineered strain for the biosynthesis of LNFP I was reported by rational design of metabolic pathways in Escherichia coli. The engineered strains produced high yields of LNFP I in fed-batch cultivation, with a conversion rate of lacto-N-tetraose to LNFP I reaching the highest level reported so far. The LNFP I-produced platform established here is broadly suitable for the production of complex and branched oligosaccharides.
CARBOHYDRATE POLYMERS
(2022)
Article
Nutrition & Dietetics
Ambra Giorgetti, Daniela Paganini, Suzane Nyilima, Robert Kottler, Max Frick, Simon Karanja, Thierry Hennet, Michael B. Zimmermann
Summary: This study aimed to test whether the intake of 3.0 g of galacto-oligosaccharides (GOS) or human milk oligosaccharides (HMO) enhances iron absorption from iron-fortified maize porridge in partially breastfed Kenyan infants, and whether the maternal HMO profile modulates these effects. The results showed that the intake of GOS increased iron absorption by 78%, while the intake of HMO did not affect iron absorption. Therefore, this study suggests that 2'-fucosyllactose (2'-FL) and lacto-N-neotetraose (LNnT) do not have a physiological role in facilitating iron absorption during infancy.
AMERICAN JOURNAL OF CLINICAL NUTRITION
(2023)
Article
Biochemistry & Molecular Biology
Chihaya Yamada, Takane Katayama, Shinya Fushinobu
Summary: Glycoside hydrolase family 136 (GH136) was established based on the discovery and structural analysis of lacto-N-biosidase (LNBase) from Bifidobacterium longum subsp. longum JCM1217 (BlLnbX). Homologous genes of BlLnbX are widely found in human gut bacteria and monkey Bifidobacterium spp. Other bacterial strains like Bifidobacterium saguini, Tyzzerella nexilis, and Ruminococcus lactaris also exhibit LNBase activity. Crystal structures of LNBases from B. saguini and T. nexilis were determined in complex with lacto-N-biose I, showing a conserved fold and catalytic residues.
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
(2022)
Article
Chemistry, Applied
Zeyu Li, Yingying Zhu, Zhaolin Huang, Pan Zhang, Wenli Zhang, Wanmeng Mu
Summary: An efficient LNFP I-producing strain without by-product 2'-FL was developed by stepwise de novo pathway construction. LNTri II-producing strains were created by integrating beta 1,3-N-acetylglucosaminyltransferase, which can be further converted to LNT by LNT-producing beta 1,3-galactosyltransferase. The de novo and salvage pathways of GDP-fucose were introduced into highly efficient LNT-producing chassis. Specific alpha 1,2-fucosyltransferase was verified to eliminate by-product 2'-FL.
CARBOHYDRATE POLYMERS
(2023)
Article
Biochemistry & Molecular Biology
Shuntaro Machida, Katsuichi Saito, Mamoru Nishimoto, Motomitsu Kitaoka
Summary: This study reports a method to produce Lacto-N-biose I (LNB) without the use of genetically modified enzymes. By selectively inactivating interfering enzymes, GlcNAc was successfully converted into LNB.
JOURNAL OF APPLIED GLYCOSCIENCE
(2022)
Article
Chemistry, Applied
Yuanyuan Liu, Aijun Tong, Xiaoxiang Gao, Sinan Yuan, Ruting Zhong, Chao Zhao
Summary: The translation abstract introduces a new enzyme, Tp2FT, which may have potential for synthesizing important fucosylated oligosaccharide LNFP I. The study found that LNFP I can reduce the concentrations of inflammatory cytokines and effectively inhibit the synthesis of enterovirus 71 proliferation, suggesting its potential therapeutic effect on hand foot mouth disease.
Article
Materials Science, Paper & Wood
Masahisa Wada, Sayuri Wakiya, Kayoko Kobayashi, Satoshi Kimura, Motomitsu Kitaoka, Ryosuke Kusumi, Fumiko Kimura, Tsunehisa Kimura
Summary: Enzymatic synthesis was used to produce cellulose oligomers with azido groups, which were then used to create cellulose microcrystal suspensions with exposed azido groups on the surface. The microcrystals, in a crystalline form of cellulose II, were several micrometers in length and had a thickness of 5.1-5.2 nm.
Article
Biochemical Research Methods
Masahiro Nakajima, Nobukiyo Tanaka, Kaito Kobayashi, Hiroyuki Nakai, Satoshi Kimura, Tadahisa Iwata, Hayao Taguchi
Summary: Beta-(1 -> 2)-glucans were synthesized using different methods to achieve varying degrees of polymerization, with beta-(1 -> 2)-glucans up to approximately 280 obtained using sucrose phosphorolysis as a source of mo-glucose 1-phosphate. A colorimetric assay of beta-glucosidase activity was used to determine the degrees of polymerization of the beta-(1 -> 2)-glucan products.
ANALYTICAL BIOCHEMISTRY
(2021)
Article
Biotechnology & Applied Microbiology
Miriam N. Ojima, Yuya Asao, Aruto Nakajima, Toshihiko Katoh, Motomitsu Kitaoka, Aina Gotoh, Junko Hirose, Tadasu Urashima, Satoru Fukiya, Atsushi Yokota, Maher Abou Hachem, Mikiyasu Sakanaka, Takane Katayama
Summary: In this study, the authors examined the HMO consumption phenotypes of two bifidobacterial species and found that the diversity of the FL transporter led to uptake specificities of fucosylated sugars ranging from simple trisaccharides to complex hexasaccharides. These findings explain the adaptation and proliferation of bifidobacteria in the competitive and HMO-rich infant gut environment.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Kazuhiro Chiku, Mitsuru Yoshida, Hiroshi Ono, Motomitsu Kitaoka
Summary: The study investigated the behavior of maltose and maltulose during heating in a phosphate buffer solution at pH 7.5, revealing that maltose was isomerized into maltulose which then underwent degradation and tautomerization to form 3-deoxy-D-glycero-pent-2-ulose. The formation of different products under varying acid-base conditions was also observed.
CARBOHYDRATE RESEARCH
(2021)
Article
Biochemistry & Molecular Biology
Toma Kashima, Kouki Okumura, Akihiro Ishiwata, Machika Kaieda, Tohru Terada, Takatoshi Arakawa, Chihaya Yamada, Kentaro Shimizu, Katsunori Tanaka, Motomitsu Kitaoka, Yukishige Ito, Kiyotaka Fujita, Shinya Fushinobu
Summary: A novel glycoside hydrolase family member, alpha-D-fructofuranosidase and difructose dianhydride I synthase/hydrolase, was identified in Bifidobacterium dentium. This enzyme has unique activities and is not homologous with known enzymes, indicating its novelty in the field of glycoside hydrolases.
JOURNAL OF BIOLOGICAL CHEMISTRY
(2021)
Article
Multidisciplinary Sciences
Wataru Saburi, Takanori Nihira, Hiroyuki Nakai, Motomitsu Kitaoka, Haruhide Mori
Summary: Glycoside phosphorylases are promising enzymes for efficient glycoside production. In this study, a new glycoside phosphorylase, solabiose phosphorylase, was characterized from an uncharacterized protein. The enzyme showed specific activity towards d-galactose, producing solabiose through a sequential bi-bi mechanism. Solabiose was successfully produced from lactose and sucrose without the need for chromatographic procedures.
SCIENTIFIC REPORTS
(2022)
Article
Biochemistry & Molecular Biology
Shuntaro Machida, Katsuichi Saito, Mamoru Nishimoto, Motomitsu Kitaoka
Summary: This study reports a method to produce Lacto-N-biose I (LNB) without the use of genetically modified enzymes. By selectively inactivating interfering enzymes, GlcNAc was successfully converted into LNB.
JOURNAL OF APPLIED GLYCOSCIENCE
(2022)
Article
Ecology
Miriam N. Ojima, Lin Jiang, Aleksandr A. Arzamasov, Keisuke Yoshida, Toshitaka Odamaki, Jinzhong Xiao, Aruto Nakajima, Motomitsu Kitaoka, Junko Hirose, Tadasu Urashima, Toshihiko Katoh, Aina Gotoh, Douwe van Sinderen, Dmitry A. Rodionov, Andrei L. Osterman, Mikiyasu Sakanaka, Takane Katayama
Summary: This study applied assembly theory to investigate the formation of bifidobacterial communities in the infant gut. The results showed that arrival order and sugar consumption phenotypes significantly affected community formation. Bifidobacterium bifidum and Bifidobacterium longum subsp. infantis dominated through inhibitory priority effects, while Bifidobacterium breve benefited from facilitative priority effects and utilized a specific degradant to dominate. This study highlights the importance of initial community assembly and its impact on the maturation trajectory of the infant gut microbiota.
Article
Biochemistry & Molecular Biology
Kazuhiro Chiku, Ayaka Ohfuji, Nozomi Ohtake, Mitsuru Yoshida, Hiroshi Ono, Motomitsu Kitaoka
Summary: Isomaltose and isomaltulose undergo isomerization and release of glucose upon heating in sodium phosphate buffer. The concentration of these disaccharides decreases with heating time, but this decrease is not directly correlated with the release of glucose or fructose. The reaction also leads to a slight decrease in pH and the formation of organic acids, along with the presence of formic acid and species resulting from hydroxyl group elimination. The released glucose is likely generated through tautomerization and cleavage of the glycosidic linkage.
CARBOHYDRATE RESEARCH
(2022)
Article
Biochemistry & Molecular Biology
Motomitsu Kitaoka
Summary: We developed a Microsoft Excel file called Enzyme Kinetics_Calculator, which contains macro programs that can automatically calculate kinetic parameters and their standard errors for typical enzymatic reactions. The file also includes functionalities to generate [S]-v plots and 1/[S]-1/v plots with theoretical lines. Enzyme Kinetics_Calculator is available as a supplementary file on the J. Appl. Glycosci. website.
JOURNAL OF APPLIED GLYCOSCIENCE
(2023)
Article
Food Science & Technology
Xin Huang, Yue Sun, Suhong Li, Motomitsu Kitaoka, Tuoping Li
Summary: To enhance the thermal stability of Lacto-N-biose (LNB), it was encapsulated within OVA-CMC microparticles. With a mass ratio of OVA:CMC:LNB as 1.5:1:1, the microparticles exhibited a particle size of 623 +/- 4 nm, encapsulation efficiency of 83 +/- 1.38%, and loading efficiency of 45 +/- 0.92%. Characterization techniques such as SEM, FT-IR, and XRD confirmed the spherical and uniform distribution of the OVA-CMC-LNB microparticles. Hydrogen bonding and electrostatic interactions were identified as the main forces responsible for the microparticles' assembly. Thermal analysis revealed that the microparticles remained structurally stable at 240 degrees C, greatly improving the thermal stability of LNB. In vitro digestion experiments demonstrated a release rate of LNB from the microparticles of 95 +/- 1.31%. These findings provide a theoretical and technological framework for enhancing the thermal stability of LNB delivery systems.
INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Junichi Mano, Hirotoshi Sushida, Tsuyoshi Tanaka, Ken Naito, Hiroshi Ono, Masakazu Ike, Ken Tokuyasu, Motomitsu Kitaoka
Summary: By extensively screening microbial strains, we discovered that the BS15 strain efficiently produces PEFAs with high titer. The strain was able to produce high concentrations of PEFA and dry cells in batch fermentation. Additionally, the genome sequence of the BS15 strain was analyzed and a high-quality reference genome was generated.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2023)
Article
Biology
Sei Motouchi, Kaito Kobayashi, Hiroyuki Nakai, Masahiro Nakajima
Summary: In this study, the structural and functional analyses of OpgG and OpgD from Escherichia coli revealed that these proteins are beta-1,2-glucanases with different activities. A reaction mechanism with a long proton transfer pathway was proposed for OpgD. The findings enhance our understanding of OPG biosynthesis and provide insights into functional diversity for this novel enzyme family.
COMMUNICATIONS BIOLOGY
(2023)
Review
Biology
Masahiro Nakajima
Summary: This review introduces enzymes associated with β-1,2-glucans, particularly glucoside phosphorylase that degrades β-1,2-glucooligosaccharides, two novel β-1,2-glucanases, and related enzymes. The review also discusses the distribution and physiological role of β-1,2-glucans in nature.
Article
Gastroenterology & Hepatology
Rika Hirano, Mikiyasu Sakanaka, Kazuto Yoshimi, Naohisa Sugimoto, Syogo Eguchi, Yuko Yamauchi, Misaki Nara, Shingo Maeda, Yuta Ami, Aina Gotoh, Takane Katayama, Noriho Iida, Tamotsu Kato, Hiroshi Ohno, Satoru Fukiya, Atsushi Yokota, Mamoru Nishimoto, Motomitsu Kitaoka, Hiroyuki Nakai, Shin Kurihara
Summary: The study identified Gal-beta 1,4-Rha as a next-generation prebiotic candidate that selectively promotes the growth of beneficial bacteria like Bifidobacterium without stimulating the growth of other prominent bacteria and pathogens. This prebiotic was found to suppress the growth of the enteric pathogen Clostridioides difficile, indicating its potential for preventing infections and improving gut health.
