DOCSLIB.ORG

A Novel Strategy for D-Psicose and Lipase Co-Production Using a Co

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
A Novel Strategy for D-Psicose and Lipase Co-Production Using a Co Zhang et al. Bioresour. Bioprocess. (2021) 8:77 https://doi.org/10.1186/s40643-021-00429-8 RESEARCH Open Access A novel strategy for D-psicose and lipase co-production using a co-culture system of engineered Bacillus subtilis and Escherichia coli and bioprocess analysis using metabolomics Jun Zhang1,2,4 , Wen Luo1, Zhiyuan Wang1, Xiaoyan Chen1,4, Pengmei Lv1* and Jingliang Xu1,3* Abstract To develop an economically feasible fermentation process, this study designed a novel bioprocess based on the co- culture of engineered Bacillus subtilis and Escherichia coli for the co-production of extracellular D-psicose and intracel- lular lipase. After optimizing the co-culture bioprocess, 11.70 g/L of D-psicose along with 16.03 U/mg of lipase was obtained; the glucose and fructose were completely utilized. Hence, the conversion rate of D-psicose reached 69.54%. Compared with mono-culture, lipase activity increased by 58.24%, and D-psicose production increased by 7.08%. In addition, the co-culture bioprocess was explored through metabolomics analysis, which included 168 carboxylic acids and derivatives, 70 organooxygen compounds, 34 diazines, 32 pyridines and derivatives, 30 benzene and substituted derivatives, and other compounds. It also could be found that the relative abundance of diferential metabolites in the co-culture system was signifcantly higher than that in the mono-culture system. Pathway analysis revealed that, tryp- tophan metabolism and β-alanine metabolism had the highest correlation and played an important role in the co- culture system; among them, tryptophan metabolism regulates protein synthesis and β-alanine metabolism, which is related to the formation of metabolic by-products. These results confrm that the co-cultivation of B. subtilis and E. coli can provide a novel idea for D-psicose and lipase biorefnery, and are benefcial for the discovery of valuable second- ary metabolites such as turanose and morusin. Highlights Co-cultivation was frstly carried out for the co-production of D-psicose and lipase. The carbon source glucose and fructose were fully utilized during co-culture. The yield of D- psicose was 11.70 g/L, and the conversion rate reached 69.54%. Self-induced expression of lipase was realized with enzyme activity of 16.03 U/mg. Metabolomics analyses were used to explore the co-culture efects of mixed strains. Keywords: D-Psicose, Lipase, Escherichia coli, Bacillus subtilis, Co-culture, Metabolomics *Correspondence: lvpm@ms.giec.ac.cn; xujl@zzu.edu.cn Introduction 1 Guangdong Provincial Key Laboratory of New and Renewable Energy Research and Development, Guangzhou Institute of Energy Conversion, D-Psicose is a rare sugar and a new type of functional Chinese Academy of Sciences, CAS Key Laboratory of Renewable Energy, sweetener with excellent physiological efects such as Guangzhou 510640, China Full list of author information is available at the end of the article anti-hyperglycemic and anti-hyperlipidemic efects © The Author(s) 2021. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http:// creat iveco mmons. org/ licen ses/ by/4. 0/. Zhang et al. Bioresour. Bioprocess. (2021) 8:77 Page 2 of 18 (Chen et al. 2019; Zhang et al. 2016). Terefore, it is mixed microorganisms with complementary advan- widely used in food, medicine, and other felds. Presently, tages (Bertrand et al. 2014; Rosero-Chasoy et al. 2021). enzymatic conversion is the most important method for Furthermore, it has a strong ability to tolerate harsh the synthesis of D-psicose, and under the catalytic action environments, such as infection, substrate, and prod- of ketose 3-epimerases, D-fructose can be isomerized uct inhibition; hence, better fermentation results can into D-psicose, which is the current hot spot of D-psicose be obtained. Currently, there are two main forms of co- synthesis (Jiang et al. 2020). However, the use of fer- culture systems (Wein et al. 2019): (a) the co-cultivation mentation methods for the cultivation of engineered of unknown microbial fora isolated from nature and (b) Bacillus subtilis, which harbors the D-tagatose 3-epime- the co-cultivation of artifcially designed microbial fora. rase (DTEase) gene, has the potential to directly produce Compared with the former, the latter system is relatively D-psicose as reported in a previous study (Zhang et al. fragile; therefore, there should be a careful selection of 2020a). Te conversion rate of the substrate can reach the combined strains. To ensure the efcient and stable 56.26%, which is a great advantage in realizing the cou- growth of the co-culture system, the population ratio pling of cell growth and D-psicose production, thereby in a reasonable range and external conditions at certain reducing the biosynthesis costs. stages, which are favored by priority populations, has Lipase is a hydrolase which acts on the ester bond with been maintained through optimized medium compo- catalytic diversity, and it is also one of the most widely nents and strengthened process regulation (Klitgord and used biocatalysts in industrial production (Melani et al. Segre 2010). B. subtilis and E. coli are Gram-positive and 2020). Geobacillus thermocatenulatus lipase 2 (GTL2) is Gram-negative bacteria, respectively. Tey have a well- a typical thermophilic enzyme with high catalytic activity mapped genetic background and are commonly used as and good thermal stability and has several applications model organisms. Under optimized co-culture condi- in pharmaceutical and organic synthesis, chiral com- tions, they can produce efective synergistic efects and pound resolution, and bioenergy; thus, it is a promising increase the yield of target products (Faust and Raes biocatalyst with great application potential (Godoy et al. 2012). Recently, indole (an aromatic heterocyclic organic 2019; Kajiwara et al. 2020). In addition, Escherichia coli compound) was produced by co-cultivation of E. coli and has a short culture cycle with high target protein levels, B. subtilis with signifcantly improved yield (Singh et al. which makes it a good host for expressing GTL2. Under 2020). the induction of expensive IPTG, GTL2 achieved solu- Te construction of an artifcial co-culture system is ble expression in E. coli, and the enzyme activity reached relatively easy and simple, but metabolic regulation is rel- 39.50 U/mg in a previous study (Zhang et al. 2020b). atively complicated. Tus, it is necessary to use metabo- However, to enhance the fermentation economy, it is lomics analysis to explore the symbiotic efects of mixed desirable to induce GTL2 expression using α-lactose. strains. In general, metabolomics is defned as the study Presently, single-bacterial fermentation of lipase may of the overall metabolite spectrum in a system (cell, tis- cause excessive production costs, whereas mixed-bac- sue, or organism) under given conditions. It is a collection terial fermentation of expensive D-psicose can reduce of small molecular metabolites with a molecular weight the fermentation costs. Tis innovation in traditional of less than 1000 in biological samples (Patti et al. 2012). fermentation not only reduces the overall cost, but also A large amount of qualitative and quantitative metabolite improves the quality of fermentation. In addition, co- data can be obtained and classifed according to specifc culture bioprocess can also efectively utilize glucose and metabolic pathways to construct metabolic networks fructose, which is of relevance for complex substrates, and infer their physiological functions to be able to more such as waste fruit and vegetable hydrolysate (Jiang et al. comprehensively analyze the dynamic metabolic reac- 2017), to achieve the goal of carbon neutrality, and the tions of organisms in response to environmental changes. full consumption of the carbon source is conducive to the Microbial metabolites, as well as the identifcation of subsequent separation of D-psicose. Furthermore, in pre- metabolic variations in diferent environments, can easily vious studies, the extracellular production of D-psicose be screened through efective analytical methods, such in recombinant B. subtilis (Zhang et al. 2020a) and the as liquid chromatography–mass spectrometry (LC–MS) soluble expression of GTL2 in engineered E. coli (Zhang based on the UHPLC–QE-MS nontargeted metabo- et al. 2020b) have been achieved. Hence, it is better to lomics approach (Cheng et al. 2020; Xia et al. 2020). explore the possibility of co-production of D-psicose and However, there is limited information about the difer- lipase using co-culture to develop a more economical fer- ential metabolites and key metabolic pathways in the mentation process. co-culture process of B. subtilis and E. coli in the exist- Based on this principle of symbiosis (Wein et al. 2019), ing literature. Hence, the screening of major diferential the co-culture of microorganisms is used to ferment metabolites and metabolic pathways using metabolomics Zhang et al. Bioresour. Bioprocess. (2021) 8:77 Page 3 of 18 analysis can better manipulate the co-cultivation process protein solution obtained was subjected to protein level of B. subtilis and E. coli, which is benefcial to the stable analysis using SDS-PAGE. operation of the co-culture system. In this study, a co-culture system of recombinant B. Lipase activity analysis subtilis and E.
Load more

Recommended publications
  • Determining the Effect of Small Doses of Fructose and Its Epimers on Glycemic Control
    Determining the Effect of Small Doses of Fructose and its Epimers on Glycemic Control by Jarvis Clyde Noronha A thesis submitted in conformity with the requirements for the degree of Master of Science Department of Nutritional Sciences University of Toronto © Copyright by Jarvis Clyde Noronha 2017 ii Determining the Effect of Small Doses of Fructose and its Epimers on Glycemic Control Jarvis Clyde Noronha Master of Science Department of Nutritional Sciences University of Toronto 2017 Abstract Given that sugars have emerged as the dominant nutrient of concern in diabetes, there is a need for the development of alternative sweeteners. To assess the role of small doses (5g, 10g) of fructose and allulose on postprandial glucose metabolism, we conducted an acute randomized controlled trial in individuals with type 2 diabetes. We found that small doses of allulose, but not fructose, modestly reduced the postprandial glycemic response to a 75g oral glucose load. To assess whether low-dose (< 50-g/day) fructose and all its epimers (allulose, tagatose and sorbose) lead to sustainable improvements in long-term glycemic control, we conducted a systematic review and meta-analysis of controlled feeding trials. The available evidence suggested that fructose and tagatose led to significant reductions in HbA1c and fasting glucose. Our findings highlight the need for long-term randomized controlled trials to confirm the viability of fructose and its epimers as alternative sweeteners. iii Table of Contents Contents Abstract ...........................................................................................................................
    [Show full text]
  • Download Author Version (PDF)
    Environmental Science: Nano CdS Nanoparticles in Soil Induce Metabolic Reprogramming in Broad Bean (Vicia faba L.) Roots and Leaves Journal: Environmental Science: Nano Manuscript ID EN-ART-08-2019-000933.R1 Article Type: Paper Page 1 of 36 Environmental Science: Nano 1 2 3 4 The rapid development of nanotechnology has raised concern regarding the 5 6 7 environmental toxicity of nanoparticles (NPs). However, little is known about the 8 9 molecular mechanisms underlying NP toxicity in plants. Understanding toxic 10 11 12 mechanisms in organisms at molecular level, especially in crop plants, is important for 13 14 their sustainable use and development. In this study, although none of the phenotypic 15 16 17 parameters of broad bean plants (photosynthetic pigments contents, biomass, and lipid 18 19 20 peroxidation) were overtly impacted in response to CdS-NPs in soil during 28 d of 21 22 exposure, metabolomics revealed marked and statistically significant alterations in the 23 24 25 metabolite profiles of plant roots and leaves. The reprogramming of antioxidant 26 27 metabolite production presumably reflected the molecular defense response of the 28 29 30 plants to CdS-NPs stress. The sensitive responses of flavone, putrescine and 31 32 33 noradrenaline in the leaves suggests the use of these compounds in legumes as 34 35 biomarkers of oxidative stress induced by the presence of CdS-NPs in soil. 36 37 38 Metabolomics might thus be a suitable approach for the early detection of soil 39 40 contamination by Cd. In the plants, the reprogramming of carbon and nitrogen 41 42 43 metabolism (including sugars, organic acids, amino acids, and N-containing 44 45 46 compounds) alleviated the toxicity of CdS-NPs, which may have been caused by free 47 48 Cd2+ ions or perhaps by a particle-specific response.
    [Show full text]
  • Electronic Supplementary Information
    Electronic Supplementary Material (ESI) for Chemical Science. This journal is © The Royal Society of Chemistry 2019 Electronic Supplementary Information Poly(ionic liquid)s as a Distinct Receptor Material to Create Highly- Integrated Sensing Platform for Efficiently Identifying a Myriad of Saccharides Wanlin Zhang, Yao Li, Yun Liang, Ning Gao, Chengcheng Liu, Shiqiang Wang, Xianpeng Yin, and Guangtao Li* *Corresponding authors: Guangtao Li (lgt@mail.tsinghua.edu.cn) S1 Contents 1. Experimental Section (Page S4-S6) Materials and Characterization (Page S4) Experimental Details (Page S4-S6) 2. Figures and Tables (Page S7-S40) Fig. S1 SEM image of silica colloidal crystal spheres and PIL inverse opal spheres. (Page S7) Fig. S2 Adsorption isotherm of PIL inverse opal. (Page S7) Fig. S3 Dynamic mechanical analysis and thermal gravimetric analysis of PIL materials. (Page S7) Fig. S4 Chemical structures of 23 saccharides. (Page S8) Fig. S5 The counteranion exchange of PIL photonic spheres from Br- to DCA. (Page S9) Fig. S6 Reflection and emission spectra of spheres for saccharides. (Page S9) Table S1 The jack-knifed classification on single-sphere array for 23 saccharides. (Page S10) Fig. S7 Lower detection concentration at 10 mM of the single-sphere array. (Page S11) Fig. S8 Lower detection concentration at 1 mM of the single-sphere array. (Page S12) Fig. S9 PIL sphere exhibiting great pH robustness within the biological pH range. (Page S12) Fig. S10 Exploring the tolerance of PIL spheres to different conditions. (Page S13) Fig. S11 Exploring the reusability of PIL spheres. (Page S14) Fig. S12 Responses of spheres to sugar alcohols. (Page S15) Fig.
    [Show full text]
  • Sugars and Plant Innate Immunity
    Journal of Experimental Botany Advance Access published May 2, 2012 Journal of Experimental Botany, Page 1 of 10 doi:10.1093/jxb/ers129 REVIEW PAPER Sugars and plant innate immunity Mohammad Reza Bolouri Moghaddam and Wim Van den Ende* KU Leuven, Laboratory of Molecular Plant Biology, Kasteelpark Arenberg 31, B-3001 Leuven, Belgium * To whom correspondence should be addressed. E-mail: wim.vandenende@bio.kuleuven.be Received 27 January 2012; Revised 25 March 2012; Accepted 30 March 2012 Abstract Sugars are involved in many metabolic and signalling pathways in plants. Sugar signals may also contribute to Downloaded from immune responses against pathogens and probably function as priming molecules leading to pathogen-associated molecular patterns (PAMP)-triggered immunity and effector-triggered immunity in plants. These putative roles also depend greatly on coordinated relationships with hormones and the light status in an intricate network. Although evidence in favour of sugar-mediated plant immunity is accumulating, more in-depth fundamental research is required to unravel the sugar signalling pathways involved. This might pave the way for the use of biodegradable http://jxb.oxfordjournals.org/ sugar-(like) compounds to counteract plant diseases as cheaper and safer alternatives for toxic agrochemicals. Key words: immunity, pathogen, priming, signal, sugar. Introduction by guest on March 24, 2016 Sugars such as glucose, fructose, and sucrose are recognized Chisholm et al., 2006). Sugars are well known to activate as signalling molecules in plants (Rolland et al., 2006; various pattern recognition genes (Herbers et al., 1996a,b; Bolouri-Moghaddam et al., 2010), in addition to their Johnson and Ryan, 1990).
    [Show full text]
  • FDA Finalizes Allulose Guidance and Requests Information on Other Sugars Metabolized Differently Than Traditional Sugars
    FDA Finalizes Allulose Guidance and Requests Information on Other Sugars Metabolized Differently Than Traditional Sugars October 19, 2020 Food, Drug, and Device FDA has taken two notable actions regarding the sugars declaration in the Nutrition Facts Label (NFL) and Supplement Facts Label (SFL). On Friday, the agency released a final guidance regarding the declaration of allulose, confirming that this monosaccharide need not be included in the declaration of “Total Sugars” or “Added Sugars,” though it must be included in the “Total Carbohydrates” declaration in the NFL. Today, FDA published a Federal Register notice requesting information about and comments on the nutrition labeling of other sugars that are metabolized differently than traditional sugars. We briefly summarize both documents below to help inform stakeholder comments on the notice, which are due to FDA by December 18, 2020. Allulose Final Guidance Allulose, or D-psicose, is a monosaccharide that can be used as a substitute for traditional sugar in food and beverage products. For purposes of nutrition labeling, FDA has generally 1 defined nutrients based on their chemical structure.0F Accordingly, when FDA updated its NFL and SFL regulations in 2016, the agency reiterated the definition of “Total Sugars” as the sum of 2 all free monosaccharides and disaccharides (e.g. glucose, fructose, and sucrose).1F FDA also added to these regulations a definition of “Added Sugars” – sugars added during the processing 3 of food, or packaged as such – and required their declaration in the NFL and SFL.2F Although the agency recognized that there are sugars that are metabolized differently than traditional sugars, FDA did not make a determination at that time as to whether allulose should be excluded from “Total Carbohydrate,” “Total Sugars,” or “Added Sugars” Declarations.
    [Show full text]
  • Effects of D-Allulose on Glucose Tolerance and Insulin Response to A
    Clinical care/Education/Nutrition Open access Original research BMJ Open Diab Res Care: first published as 10.1136/bmjdrc-2020-001939 on 26 February 2021. Downloaded from Effects of D- allulose on glucose tolerance and insulin response to a standard oral sucrose load: results of a prospective, randomized, crossover study Francesco Franchi ,1 Dmitry M Yaranov,1 Fabiana Rollini,1 Andrea Rivas,1 Jose Rivas Rios,1 Latonya Been,1 Yuma Tani,2 Masaaki Tokuda,3 Tetsuo Iida,2 Noriko Hayashi,2 Dominick J Angiolillo,1 Arshag D Mooradian1 To cite: Franchi F, Yaranov DM, ABSTRACT Rollini F, et al. Effects of D- Introduction Current dietary guidelines recommend Significance of this study allulose on glucose tolerance limiting sugar intake for the prevention of diabetes and insulin response to a mellitus (DM). Reduction in sugar intake may require sugar What is already known about this subject? standard oral sucrose load: substitutes. Among these, D- allulose is a non- calorie rare ► D- allulose is defined one of the rare sugars, results of a prospective, which has been shown in animal and clinical randomized, crossover study. monosaccharide with 70% sweetness of sucrose, which has shown anti- DM effects in Asian populations. However, studies, conducted mostly in Asian populations, BMJ Open Diab Res Care to have postprandial plasma glucose suppres- 2021;9:e001939. doi:10.1136/ there is limited data on the effects of D- allulose in other sive effects with antiobesity and antidiabetic bmjdrc-2020-001939 populations, including Westerners. Research design and methods This was a prospective, effects. copyright. randomized, double- blind, placebo- controlled, crossover What are the new findings? ► Supplemental material is study conducted in 30 subjects without DM.
    [Show full text]
  • Sweetness and Sensory Properties of Commercial and Novel Oligosaccharides Of
    1 Sweetness and sensory properties of commercial and novel oligosaccharides of 2 prebiotic potential 3 4 Laura Ruiz-Aceitunoa, Oswaldo Hernandez-Hernandeza, Sofia Kolidab, F. Javier 5 Morenoa,* and Lisa Methvenc 6 7 a Institute of Food Science Research, CIAL (CSIC-UAM), Nicolás Cabrera 9, 28049 8 Madrid (Spain) 9 b OptiBiotix Health plc, Innovation Centre, Innovation Way, Heslington, York YO10 10 5DG (UK) 11 c Sensory Science Centre, Department of Food and Nutritional Sciences, The University 12 of Reading, PO Box 226, Whiteknights, Reading RG6 6AP (UK) 13 14 *Corresponding author: javier.moreno@csic.es Tel (+34) 91 0017948 1 15 Abstract 16 This study investigates the sweetness properties and other sensory attributes of ten 17 commercial and four novel prebiotics (4-galactosyl-kojibiose, lactulosucrose, lactosyl- 18 oligofructosides and raffinosyl-oligofructosides) of high degree of purity and assesses the 19 influence of their chemical structure features on sweetness. The impact of the type of 20 glycosidic linkage by testing four sucrose isomers, as well as the monomer composition 21 and degree of polymerization on sweetness properties were determined. Data from the 22 sensory panel combined with principal component analysis (PCA) concludes that chain 23 length was the most relevant factor in determining the sweetness potential of a 24 carbohydrate. Thus, disaccharides had higher sweetness values than trisaccharides which, 25 in turn, exhibited superior sweetness than mixtures of oligosaccharides having DP above 26 3. Furthermore, a weak non-significant trend indicated that the presence of a ketose sugar 27 moiety led to higher sweetness. The novel prebiotics tested in this study had between 18 28 and 25% of relative sweetness, in line with other commercial prebiotics, and samples 29 varied in their extent of off flavour.
    [Show full text]
  • Author Correction: the Planctomycete Stieleria Maiorica Mal15t Employs Stieleriacines to Alter the Species Composition in Marine Biofilms
    https://doi.org/10.1038/s42003-020-01271-y OPEN Author Correction: The planctomycete Stieleria maiorica Mal15T employs stieleriacines to alter the species composition in marine biofilms Nicolai Kallscheuer , Olga Jeske, Birthe Sandargo, Christian Boedeker, Sandra Wiegand , Pascal Bartling, 1234567890():,; Mareike Jogler, Manfred Rohde, Jörn Petersen, Marnix H. Medema , Frank Surup & Christian Jogler Correction to: Communications Biology https://doi.org/10.1038/s42003-020-0993-2, published online 12 June 2020. In the original version of the Article “The planctomycete Stieleria maiorica Mal15T employs stieleriacines to alter the species composition in marine biofilms”, the authors described the new genus Stieleria and its type species Stieleria maiorica. However, the descriptions were not presented in the paper in the format required by Rule 27 and Rule 30 of the International Code of Nomenclature of Prokaryotes. Therefore, we here present the descriptions in the correct format. Description of Stieleria gen. nov. Stieleria (Stie.le’ri.a. N.L. fem. n. Stieleria named in honor of Anja Heuer, née Stieler, an extraordinary skilled German technician at the Leibniz Institute DSMZ, who played a key role in the cultivation of literally hundreds of novel planctomycetal strains). The round-to- pear-shaped cells with a smooth cell surface form rosettes or short chains. Cells reproduce by polar budding. In liquid culture, they produce an extracellular matrix that interconnects cells in aggregates. Daughter cells are motile, while mother cells are non-motile. The lifestyle is heterotrophic, obligatory aerobic, and mesophilic. The genus belongs to the phylum Planctomycetes, class Planctomycetia, order Pirellulales, and family Pirellulaceae. The type species is Stieleria maiorica.
    [Show full text]
  • Characterization of EPS-Producing Brewery-Associated Lactobacilli
    TECHNISCHE UNIVERSITÄT MÜNCHEN Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt Lehrstuhl für Technische Mikrobiologie Characterization of EPS-producing brewery-associated lactobacilli Marion Elisabeth Fraunhofer Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt der Technischen Universität München zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften genehmigten Dissertation. Vorsitzender: Prof. Dr. Horst-Christian Langowski Prüfer der Dissertation: 1. Prof. Dr. Rudi F. Vogel 2. Hon.-Prof. Dr.-Ing. Friedrich Jacob Die Dissertation wurde am 13.12.2017 bei der Technischen Universität München eingereicht und durch die Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt am 06.03.2018 angenommen. Vorwort Vorwort Die vorliegende Arbeit entstand im Rahmen eines durch Haushaltsmittel des BMWi über die AiF-Forschungsvereiningung Wissenschaftsförderung der Deutschen Brauwirtschaft e.V. geförderten Projekts (AiF18194 N). Mein besonderer Dank gilt meinem Doktorvater Herrn Prof. Dr. rer. nat. Rudi F. Vogel für das Vertrauen, die Unterstützung und die Loyalität auf die ich mich in den letzten Jahren stets verlassen konnte. Danke Rudi, dass ich meine Promotion an deinem Lehrstuhl durchführen durfte. Desweitern bedanke ich mich bei Dr. rer. nat. Frank Jakob, der mich durch seine umfassende Betreuung sicher durch meine Dissertation begleitet hat. Apl. Prof. Dr. Matthias Ehrmann und apl. Prof. Dr. Ludwig Niessen
    [Show full text]
  • Determination of Carbohydrates in Honey Manali Aggrawal, Jingli Hu and Jeff Rohrer, Thermo Fisher Scientific, Sunnyvale, CA
    Determination of carbohydrates in honey Manali Aggrawal, Jingli Hu and Jeff Rohrer, Thermo Fisher Scientific, Sunnyvale, CA ABSTRACT RESULTS SAMPLE ANALYSIS METHOD ACCURACY Table 7. Adulteration parameters for HS6 adulterated with 10% SS1 through SS5. Purpose: To develop an HPAE-PAD method for the determination of carbohydrates in honey Honey sugar analysis Sample Recovery HS6 (Wild Mountain Honey) samples to evaluate their quality and to assess the possibility of adulteration. Separation Adulteration Honey sugars were separated using a Dionex CarboPac PA210-Fast-4μm column (150 × 4 mm) in Method accuracy was evaluated by measuring recoveries of 10 sugar standards spiked into honey Parameters 100% + 10% + 10% + 10% + 10% + 10% For this study, we purchased 12 commercial honey samples (Table 1) and analyzed them using Honey SS1 SS2 SS3 SS4 SS5 Methods: Separation of individual honey sugars was achieved on the recently introduced Thermo series with a Dionex CarboPac PA210 guard column (50 × 4 mm). The column selectivity allow samples. For spiking experiments, four honey samples were used (HS7–HS10) and spiked with a 10- HPAE-PAD. Figure 3 shows the representative chromatograms of 3 honey samples. For all 12 Glucose(G), mg/L 121 115 116 117 119 107 Scientific™ Dionex™ CarboPac™ PA210-Fast-4μm column. Carbohydrate detection was by pulsed carbohydrates to be separated with only a hydroxide eluent generated using an eluent generator. A sugar standard mix at two concentration levels. Figure 4 shows the representative chromatograms investigated honey samples, fructose and glucose (Peak 2 and Peak 3), were found to be the major Fructose(F), mg/L 127 115 115 116 126 116 amperometric detection (PAD) with a gold working electrode and, therefore, no sample derivatization solution of honey sugar standards was prepared and an aliquot (10 μL) of the solution was injected of unspiked and spiked honey sample HS7.
    [Show full text]
  • The Food Lawyers® Respectfully Request That FDA Implements the Following
    December 7, 2020 Dockets Management Staff (HFA-305) Filed Electronically Food and Drug Administration https://www.regulations.gov Re: Sugars Metabolized Differently than Traditional Sugars (FDA-2020-N-1359) Ladies and Gentlemen: One Page Executive Summary FDA’s seeks information to “… promote the public health and help consumers make informed dietary decisions” regarding sugars that are metabolized differently than traditional sugars. Given the nation’s battles with diabetes and obesity, and the benefits that non-traditional sugars can offer in these battles, the Agency’s stated public policy goal goes to the very heart of American consumers’ health. This laudatory public policy’s realization is complicated by a lack of consumer awareness of how some sugars are metabolized differently than others. In an effort to answer the questions posed by the Agency regarding the treatment of Sugars that Are Metabolized Differently Than Traditional Sugars, we suggest that the Agency adapt a mechanism that will seek to harmonize the public policy of promoting public health with consumers’ lack of awareness of sugars that are metabolized differently than sucrose. In particular, we suggest that FDA should consider the following: 1. Establish a new category of sugars called Rare Sugars that exhibit the following characteristics: a. Are naturally occurring b. Impart a sweet taste that is at least 50% the sweetness of sucrose c. 2.0 kcal/g or less. d. Resulting pH of 6.0 or greater of dental plaque after consumption. e. No or low glycemic response. f. No or low insulinemic response. 2. Exclude Rare Sugars from “Total Sugars” and “Added Sugars” declarations to stimulate their deployment by industry and consumption by the public.
    [Show full text]
  • Natural Alternative Sweeteners and Diabetes Management
    Current Diabetes Reports (2019) 19:142 https://doi.org/10.1007/s11892-019-1273-8 LIFESTYLE MANAGEMENT TO REDUCE DIABETES/CARDIOVASCULAR RISK (B CONWAY AND H KEENAN, SECTION EDITORS) Natural Alternative Sweeteners and Diabetes Management Emily Mejia1 & Michelle Pearlman2 # Springer Science+Business Media, LLC, part of Springer Nature 2019 Abstract Purpose of Review The goal of this review is to discuss the data on natural alternative sweeteners and their effects on glucose homeostasis and other metabolic parameters within the past five years. We sought to answer whether common natural alternative sweeteners have a positive or negative effect on glucose control in both human and animal models, and whether the data supports their widespread use as a tool to help reduce the prevalence of diabetes and associated comorbid conditions. Recent Findings Recent studies suggest that natural alternative sweeteners may reduce hyperglycemia, improve lipid metabo- lism, and have antioxidant effects particularly in those that have baseline diabetes. Summary Diabetes and metabolic syndrome have become a global healthcare crisis and the sugar overconsumption plays a major role. The use of artificial sweeteners has become more prevalent to improve insulin resistance in those with diabetes, obesity, and metabolic syndrome, although the evidence does not support this result. There are however some promising data to suggest that natural alternative sweeteners may be a better alternative to sugar and artificial sweeteners. Keywords Natural alternative sweeteners . Diabetes . Stevia . Sugar alcohols . Rare sugars Introduction retinopathy, nephropathy, cerebrovascular disease, peripheral vascular disease, and cardiovascular disease [2–4]. Type 2 Diabetes mellitus (DM) is a metabolic disorder that results DM (T2DM) often coexists with other metabolic disorders from glucose dysregulation.
    [Show full text]