<Emphasis Type="Italic">In vitro</Emphasis> colonization of hydrophilic contact lenses by <Emphasis Type="Italic">Aspergillus niger</Emphasis>

In vitro colonization of hydrophilic contact lenses by Aspergillus niger was investigated. Five strains of the fungus, four polymers, two culture media and four incubation periods were considered for analysis. Only the 2700 strain colonized the lenses. The degrees of adhesion and invasion varied significantly according to the characteristics of the culture under investigation. Journal of Industrial Microbiology & Biotechnology (2002) 29, 6–9 doi:10.1038/sj.jim.7000255 Received 06 August 2001/ Accepted in revised form 23 March 2002     

Ethanol production from corn cob hydrolysates by <Emphasis Type="Italic">Escherichia coli</Emphasis> KO11

Corn cob hydrolysates, with xylose as the dominant sugar, were fermented to ethanol by recombinant Escherichia coli KO11. When inoculum was grown on LB medium containing glucose, fermentation of the hydrolysate was completed in 163 h and ethanol yield was 0.50 g ethanol/g sugar. When inoculum was grown on xylose, ethanol yield dropped, but fermentation was faster (113 h). Hydrolysate containing 72.0 g/l xylose and supplemented with 20.0 g/l rice bran was readily fermented, producing 36.0 g/l ethanol within 70 h. Maximum ethanol concentrations were not higher for fermentations using higher cellular concentration inocula. A simulation of an industrial process integrating pentose fermentation by E. coli and hexose fermentation by yeast was carried out. At the first step, E. coli fermented the hydrolysate containing 85.0 g/l xylose, producing 40.0 g/l ethanol in 94 h. Baker's yeast and sucrose (150.0 g/l) were then added to the spent fermentation broth. After 8 h of yeast fermentation, the ethanol concentration reached 104.0 g/l. This two-stage fermentation can render the bioconversion of lignocellulose to ethanol more attractive due to increased final alcohol concentration. Journal of Industrial Microbiology & Biotechnology (2002) 29, 124–128 doi:10.1038/sj.jim.7000287 Received 20 February 2002/ Accepted in revised form 04 June 2002     

Isolation of a mutant strain of Pseudomonas sp ATCC 31461 exhibiting elevated polysaccharide production

A mutant strain of the bacterium Pseudomonas sp. ATCC 31461 that exhibited elevated production of the polysaccharide gellan on glucose or corn syrup as a carbon source was isolated. Gellan production by the mutant strain was about twofold higher than its parent strain on glucose or corn syrup after 48 h of growth, and about 1.4-fold higher after 72 h. An increase in biomass production was not correlated with enhanced gellan synthesis by the mutant strain. The increased gellan production by the mutant strain on either carbon source resulted in an increase in its culture medium viscosity and the viscosity of the isolated polysaccharide produced by glucose-grown cells. No differences in the glucuronic acid content of the polysaccharides produced by the mutant and parent strains were observed. Journal of Industrial Microbiology & Biotechnology (2002) 29, 185–188 doi:10.1038/sj.jim.7000278 Received 13 February 2002/ Accepted in revised form 20 May 2002     

Influence of straw types and nitrogen sources on mushroom composting emissions and compost productivity

The effects of different straw types and organic and inorganic nitrogen (N) sources on the chemical composition and odor concentration (OC) of mushroom composting emissions, compost parameters, and mushroom yield were examined using bench-scale and large-scale (windrows and aerated tunnels) composting systems. There were close correlations between the butanol or combined H₂S+dimethyl sulfide (DMS) concentration and OC of air samples taken from different composting ingredients (r=0.83 and 0.76–0.87, P<0.01, for log_e-transformed data). Differences in N availability, in terms of NH₃ and N losses during composting, were found between different N sources. Materials in which the N was less available (chipboard and digester wastes, cocoa shells, ammonium sulfate) produced lower mushroom yields than materials in which the N was more readily available (poultry manure, urea, brewers' grains, hop and molasses wastes, cocoa meal). Replacement of poultry manure with the other N sources at 50–100% or wheat straw with rape, bean, or linseed straw in aerated tunnel or windrow composts reduced the OC and emissions of odorous sulfur-containing compounds, but also reduced yield. Urea and cocoa meal may be suitable for “low odor” prewetting of straw, with addition of poultry manure immediately before aerated tunnel composting. Rape straw in compost reduces the formation of anaerobic zones and resulting odorous emissions, since it maintains its structure and porosity better than wheat straw. Journal of Industrial Microbiology & Biotechnology (2002) 29, 99–110 doi:10.1038/sj.jim.7000292 Received 08 January 2002/ Accepted in revised form 20 June 2002     

Large-scale production of GDP-fucose and Lewis X by bacterial coupling

A large-scale production system of GDP-fucose (GDP-Fuc) and fucosylated oligosaccharides was established by the combination of recombinant Escherichia coli cells overexpressing GDP-Fuc biosynthetic genes and Corynebacterium ammoniagenes cells. E. coli cells overexpressed the genes for glucokinase, phosphomannomutase, mannose-1-phosphate guanylyltransferase, GDP-mannose (GDP-Man) dehydratase, and GDP-4-keto-6-deoxy-mannose (GKDM) epimerase/reductase as well as phosphoglucomutase and phosphofructokinase. C. ammoniagenes contributed to the formation of GTP from GMP. GDP-Fuc accumulated to 29 mM (18.4 g l⁻¹) after a 22-h reaction starting with GMP and mannose through introducing the two-step reaction to overcome the inhibition of GDP-Fuc on GDP-Man dehydratase activity. When E. coli cells overexpressing the α1,3-fucosyltransferase gene of Helicobacter pylori were put into the GDP-Fuc production system, Lewis X [Galβ1–4(Fucα1–3)GlcNAc] was produced at an amount of 40 mM (21 g l⁻¹) for 30 h from GMP, mannose, and N-acetyl lactosamine. The production system through bacterial coupling can be applied to the industrial manufacture of fucosylated oligosaccharides. Journal of Industrial Microbiology & Biotechnology (2000) 25, 213–217. Received 01 May 2000/ Accepted in revised form 20 July 2000     

Planktonic nitrate-reducing bacteria and sulfate-reducing bacteria in some western Canadian oil field waters

Oil fields that use water flooding to enhance oil recovery may become sour because of the production of H₂S from the reduction of sulfate by sulfate-reducing bacteria (SRB). The addition of nitrate to produced waters can stimulate the activities of nitrate-reducing bacteria (NRB) and control sulfide production. Many previous studies have focused on chemolithotrophic bacteria that can use thiosulfate or sulfide as energy sources while reducing nitrate. Little attention has been given to heterotrophic NRB in oil field waters. Three different media were used in this study to enumerate various types of planktonic NRB present in waters from five oil fields in western Canada. The numbers of planktonic SRB and bacteria capable of growth under aerobic conditions were also determined. In general, microbial numbers in the produced waters were very low (<10 ml⁻¹) in samples taken near or at wellheads. However, the numbers increased in the aboveground facilities. No thiosulfate-oxidizing NRB were detected in the oil field waters, but other types of NRB were detected in 16 of 18 produced water samples. The numbers of heterotrophic NRB were equal to or greater than the number of sulfide-oxidizing, chemolithotrophic NRB in 12 of 15 samples. These results showed that each of the oil fields contained NRB, which might be stimulated by nitrate amendment to control H₂S production by SRB. Journal of Industrial Microbiology & Biotechnology (2002) 29, 83–92 doi:10.1038/sj.jim.7000274 Received 20 February 2002/ Accepted in revised form 14 May 2002     

Production of butanol from starch-based waste packing peanuts and agricultural waste

We examined the fermentation of starch-based packing peanuts and agricultural wastes as a source of fermentable carbohydrates using Clostridium beijerinckii BA101. Using semidefined P2 medium containing packing peanuts and agricultural wastes, instead of glucose as a carbohydrate source, we measured characteristics of the fermentation including solvent production, productivity, and yield. With starch as substrate (control), the culture produced 24.7 g l⁻¹ acetone–butanol–ethanol (ABE), while with packing peanuts it produced 21.7 g l⁻¹ total ABE with a productivity of 0.20 g l⁻¹ h⁻¹ and a solvent (ABE) yield of 0.37. Cell growth in starch, packing peanuts, and agricultural wastes medium was different, possibly due to the different nature of these substrates. Using model agricultural waste, 20.3g l⁻¹ ABE was produced; when using actual waste, 14.8 g l⁻¹ ABE was produced. The use of inexpensive substrates will increase the economic viability of the conversion of biomass to butanol, and can provide new markets for these waste streams. Journal of Industrial Microbiology & Biotechnology (2002) 29, 117–123 doi: 10.1038/sj.jim.7000285 Received 14 November 2001/ Accepted in revised form 07 June 2002     

Synthesis and transformations of a pyrazole containing <Emphasis Type="Italic">α</Emphasis>,<Emphasis Type="Italic">β</Emphasis>-didehydro-<Emphasis Type="Italic">α</Emphasis>-amino acid derivatives

Summary.  2H-Pyran-2-ones 1 were transformed with various hydrazines into (E)- or (Z)-α,β-didehydro-α-amino acid (DDAA) derivatives 4 (and 7) containing a highly substituted pyrazolyl moiety attached at the β-position. With heterocyclic hydrazines, the products 4 were accompanied also by decarboxylated enamines E-6. In order to separate (E/Z)-mixtures of acids, they were transformed to the corresponding methyl esters 9 and 10 by the application of diazomethane. Catalytic hydrogenation under high pressures with Pd/C as a catalyst resulted in the formation of racemic alanine derivatives 11. Received January 29, 2002 Accepted May 27, 2002 Published online December 18, 2002 RID="*" ID="*"  Dedicated with deep respect to Professor Waldemar Adam on the occasion of his 65^th birthday. Acknowledgements We thank the Ministry of Education, Science and Sport of the Republic of Slovenia for the financial support (P0-0503-103). Dr. B. Kralj and Dr. D. Žigon (Center for Mass Spectroscopy, “Jožef Stefan” Institute, Ljubljana, Slovenia) are gratefully acknowledged for the mass measurements. Authors' address: Prof. Marijan Kočevar, Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, SI-1000 Ljubljana, Slovenia, E-mail: marijan.kocevar@uni-lj.si     

Monovalent cation transporters; establishing a link between bioinformatics and physiology

Toxic aluminum (Al) ion is a major constraint to plant growth in acid soils. Aluminum tolerance in wheat (Triticum aestivum L.) is strongly related to the Al-triggered efflux of malate from root apices. A role of the secreted malate has been postulated to be in chelating Al and thus excluding it from root apices (malate hypothesis), but the actual process has yet to be fully elucidated. We measured Al content and root growth during and after Al exposure using seedlings of near-isogenic lines [ET8 (Al tolerant) and ES8 (Al sensitive)] differing in the capacity to induce Al-triggered malate efflux. Aluminum doses that caused 50% root growth inhibition during 24-h exposure to Al in calcium (Ca) solution (0.5 mM CaCl₂, pH 4.5) were 50 μM in ET8 and 5 μM in ES8. Under such conditions, the amount of Al accumulated in root apices was approximately 2-fold higher in ET8 than ES8. Al-treated seedlings were then transferred to the Al-free Ca solution for 24 h. Compared to control roots (no Al pretreatment), root regrowth of Al-treated roots was about 100% in ET8 and about 25% in ES8. The impaired regrowth in ES8 was observed even after 24-h exposure to 2.5 μM Al which had caused only 20% root growth inhibition. The addition of malate (100 μM) during exposure to 50 μM Al in ES8 enhanced root growth 1.6 times and regrowth in Al-free solution 7 times, resulting in similar root growth and regrowth as in ET8. Short-term Al treatments of ES8 for up to 5 h indicated that the Al-caused inhibition of root regrowth started after 1-h exposure to Al. The stimulating effect of malate on root regrowth was observed when malate was present during Al exposure, but not when roots previously exposed to Al were rinsed with malate, although Al accumulation in root apices was similar under these malate treatments. We conclude that the malate secreted from root apices under Al exposure is essential for the apices to commence regrowth in Al-free medium, the trait that is not related to the exclusion of Al from the apices.     

Structural analysis of the curdlan-like exopolysaccharide produced by Cellulomonas flavigena KU

Cellulomonas flavigena KU produces large quantities of an insoluble exopolysaccharide (EPS) under certain growth conditions. The EPS has previously been shown to be a glucose polymer and to have solubility properties similar to curdlan, a β-1,3-D-glucan produced by Alcaligenes faecalis var. myxogenes 10C3K. Furthermore, EPS purified by alkaline extraction stains with aniline blue, a dye specific for curdlan-type polysaccharides. However, EPS-producing colonies of C. flavigena KU do not stain on aniline blue agar as do those of curdlan-producing bacteria. These facts prompted a more thorough structural analysis of the EPS. Here we report that purified EPS is indeed identical to curdlan in primary structure, but that the native form of the EPS may differ from curdlan in physical conformation. Journal of Industrial Microbiology & Biotechnology (2002) 29, 200–203 doi:10.1038/sj.jim.7000277 Received 19 February 2002/ Accepted in revised form 20 May 2002     

Evaluating the potential efficacy of three antifungal sealants of duct liner and galvanized steel as used in HVAC systems

Current recommendations for remediation of fiberglass duct materials contaminated with fungi specify complete removal, which can be extremely expensive, but in-place duct cleaning may not provide adequate protection from regrowth of fungal contamination. Therefore, a common practice in the duct-cleaning industry is the postcleaning use of antifungal surface coatings with the implication that they may contain or limit regrowth. However, even the proper use of these products has generally been discouraged because little research has been conducted on the effectiveness of most products as used in heating, ventilating, and air-conditioning (HVAC) systems. Three different coatings were evaluated on fiberglass duct liner (FGDL). Two of the three coatings were able to limit growth in the 3-month study; the third did not. One of the coatings that was able to limit growth was further evaluated in a comparison of FGDL or galvanized steel (GS) under conditions that mimicked their use in HVAC systems. The results showed that both moderately soiled and heavily soiled uncoated FGDL and GS duct material can support fungal growth, but that GS duct material was more readily cleaned. The use of an antifungal coating helped limit, but did not fully contain, regrowth on FGDL. No regrowth was detected on the coated GS. Journal of Industrial Microbiology & Biotechnology (2002) 29, 38–43 doi:10.1038/sj.jim.7000261 Received 19 November 2001/ Accepted in revised form 29 March 2002     

Use of degenerate primers and touchdown PCR to amplify a halogenase gene fragment from <Emphasis Type="Italic">Streptomyces venezuelae</Emphasis> ISP5230

Consensus amino acid sequences of FADH₂-dependent bacterial halogenases were used to design PCR primers amplifying a halogenase gene fragment from the chloramphenicol producer Streptomyces venezuelae ISP5230. The sequence-specific degenerate primers (MPF1 and MPR2) were used with a touchdown PCR procedure in the first PCR-assisted cloning of a halogenase gene fragment. In the region of the 290-bp PCR product containing the reverse primer, the deduced amino acid sequence exhibited characteristics of a β–α–β fold present in FAD-binding sites of certain monooxygenases. When used to probe Southern blots of restriction-enzyme-digested DNA, the [α-³²P]dCTP-labeled PCR product hybridized specifically with DNA fragments from genomic DNA of S. venezuelae ISP5230. Primers MPF1 and MPR2 also allowed amplification by PCR of approximately 290-bp DNA fragments from several other streptomycetes. The fragments from Streptomyces aureofaciens NRRL2209 and Streptomyces coelicolor A3(2) showed sequence identity with halogenase genes from these species. Thus, the PCR primers are of potential value for amplification and subsequent isolation of actinomycete halogenase genes. Journal of Industrial Microbiology & Biotechnology (2002) 29, 1–5 doi:10.1038/sj.jim.7000263 Received 25 June 2001/ Accepted in revised form 02 April 2002     

Synthesis of orthogonally protected (<Emphasis Type="Italic">3R</Emphasis>,<Emphasis Type="Italic">4S</Emphasis>)- and (<Emphasis Type="Italic">3S</Emphasis>,<Emphasis Type="Italic">4S</Emphasis>)-4,5-diamino-3-hydroxypentanoic acids

Summary.  The paper describes two methods of the synthesis of ethyl (3R,4S)- and (3S,4S)-4-[(benzyloxycarbonyl)amino]-5-[(tert-butyloxycarbonyl)amino]-3-hydroxypentanoates, useful for the syntheses of edeine analogs. Differently N-protected (S)-2,3-diaminopropanoic acid was used as a substrate in both procedures. The absolute configuration of newly generated asymmetric carbon atoms C-3 in β-hydroxy-γ,δ-diamino products was assigned by means of ¹H NMR spectroscopy after their transformation into corresponding piperidin-2-ones. Received May 24, 2002 Accepted October 10, 2002 Published online December 18, 2002 Acknowledgment The authors are indebted to the Faculty of Chemistry, Technical University of Gdańsk for financial support. Authors' address: Zbigniew Czajgucki, M. Sc., Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Technical University of Gdańsk, 11/12 Narutowicza St., 80-952 Gdańsk, Poland, Fax +48 58 347 11 44, E-mail: zmczaj@wp.pl     

Extracellular production of L-ascorbic acid by Chlorella protothecoides,Prototheca species,and mutants of P. moriformis during aerobic culturing at low pH

Nine strains of Chlorella protothecoides and 43 strains representing the five species of Prototheca were screened in flask culture for their ability to synthesize L-ascorbic acid (AA). Ascorbic acid was detected in all strains, ranging from 4.8 to 0.38 mg AA g⁻¹ of dry cells. Organisms selected for further study grew well and maintained their AA productivity above a pH of 3.5. They can produce AA using a variety of carbon and nitrogen sources. Aerobic fermentation of selected strains resulted in extracellular accumulation of AA up to 76 mg l⁻¹. By classical mutagenesis and selection methods, we created mutants of Prototheca moriformis ATCC 75669 that produced greater quantities of AA than the wild-type strain (78.4 vs 21.9 mg AA g⁻¹ of cells). A process based on extracellular production could greatly reduce the cost of AA manufacture by eliminating the need for extraction of the AA from the cells. Journal of Industrial Microbiology & Biotechnology (2002) 29, 93–98 doi:10.1038/sj.jim.7000275 Received 04 December 2001/ Accepted in revised form 09 May 2002     

Amylase hyperproduction by deregulated mutants of the thermophilic fungus Thermomyces lanuginosus

Thermomyces lanuginosus was subjected to three cycles of mutagenesis (UV/NTG) and a selection procedure to develop amylase-hyperproducing, catabolite-repression-resistant and partially constitutive strains. One of the selected derepressed mutant strain III₅₁, produced ∼7- and 3-fold higher specific activity of α-amylase (190 U/mg protein) and glucoamylase (105 U/mg protein), respectively, compared to a wild-type parental strain. Further, the effect of production parameters on mutant strain III₅₁ was studied using a Box–Behnken design. The regression models computed showed significantly high R ² values of 96 and 97% for α-amylase and glucoamylase activities, respectively, indicating that they are appropriate for predicting relationships between corn flour, soybean meal and pH with α-amylase and glucoamylase production. Journal of Industrial Microbiology & Biotechnology (2002) 29, 70–74 doi:10.1038/sj.jim.7000270 Received 05 July 2001/ Accepted in revised form 16 April 2002     

Chlamydospore formation of <Emphasis Type="Italic">Entoloma clypeatum</Emphasis> f. <Emphasis Type="Italic">hybridum </Emphasis>on mycorrhizas and rhizomorphs associated with <Emphasis Type="Italic">Rosa multiflora</Emphasis>

 Chlamydospores of Entoloma clypeatum f. hybridum were described on the mycorrhizas and rhizomorphs associated with Rosa multiflora. Their developmental pattern seems to be the Nyctalis type. This is the first report on chlamydospore formation on the mycorrhizae in entolomatoid fungi. Received: January 17, 2002 / Accepted: November 5, 2002 Acknowledgments K.H. is grateful to Emeritus Professor N. Sagara in Kyoto University, in whose laboratory part of this study was undertaken. Thanks are due to Mr. D. Sakuma for allowing the specimens to be kept in Osaka Museum of Natural History. Correspondence to:H. Kobayashi     

Influence of Saccharomyces cerevisiae strains on fermentation and flavor compounds of white wines made from cv. Emir grown in Central Anatolia,Turkey

The effect of inoculation with selected Saccharomyces cerevisiae strains was studied on fermentation and flavor compounds of wines made from Vitis vinifera L. cv. Emir grown in Central Anatolia, Turkey. Flavor compounds were analysed and identified by GC-FID and GC-MS, respectively. The total concentrations of flavor compounds did not increase with the addition of indigenous and commercial wine yeasts, but differences were noted in individual volatile compounds. Cluster and factor analyses of flavor compounds also showed that wines produced were different depending on the wine strain used. Wines were completely fermented to less than 1.4 g/l residual sugar. Yeasts other than S. cerevisiae survived longer than previously reported. Inoculation with selected strains increased the ethanol level. Journal of Industrial Microbiology & Biotechnology (2002) 29, 28–33 doi:10.1038/sj.jim.7000258 Received 11 July 2001/ Accepted in revised form 27 March 2002     

Optimisation of the expression of a Trametes versicolor laccase gene in Pichia pastoris

A cDNA encoding a laccase enzyme was isolated from a Trametes versicolor cDNA library. The gene was subcloned into the Pichia pastoris expression vector pPIC3.5 and transformed into the P. pastoris strains KM71 and GS115. Laccase-secreting transformants were selected by their ability to oxidise the substrate ABTS. No difference in laccase activity was observed between culture supernatants from GS115 (proteolytic) and KM71 (nonproteolytic) strains. The presence of at least 200 μM copper was necessary for optimal laccase activity in the culture supernatants. During growth of P. pastoris on minimal medium the pH of the medium was reduced to <3.0. If alanine was added to the medium the pH reduction was not as pronounced and at alanine concentrations >0.6% w/v the pH was kept constant for >7 days. Cultures in which the pH was maintained by alanine metabolism produced higher levels of laccase activity than those grown in the absence of alanine. This study describes the development of a medium that allows convenient pH control of P. pastoris without the need for continuous neutralisation. Journal of Industrial Microbiology & Biotechnology (2002) 29, 55–59 doi:10.1038/sj.jim.7000268 Received 08 August 2001/ Accepted in revised form 18 April 2002     

Gellan gum biosynthesis in Sphingomonas paucimobilis ATCC 31461: genes,enzymes and exopolysaccharide production engineering

The commercial gelling agent, gellan, is an extracellular polysaccharide (EPS) produced by Sphingomonas paucimobilis ATCC 31461. In recent years, significant progress in understanding the relationship between gellan structure and properties and elucidation of the biosynthesis and engineering of this recent product of biotechnology has been made. This review focuses on recent advances in this field. Emphasis is given to identification and characterization of genes and enzymes involved, or predicted to be involved, in the gellan biosynthetic pathway, at the level of synthesis of sugar-activated precursors, of the repeat unit assembly and of gellan polymerization and export. Identification of several genes, biochemical characterization of the encoded enzymes and elucidation of crucial steps of the gellan pathway indicate that possibilities now exist for exerting control over gellan production at any of the three levels of its biosynthesis. However, a better knowledge of the poorly understood steps and of the bottlenecks and regulation of the pathway, the characterization of the composition, structure and functional properties of gellan-like polymers produced either by the industrial strain under different culture conditions or by mutants are still required for eventual success of the metabolic engineering of gellan production. Journal of Industrial Microbiology & Biotechnology (2002) 29, 170–176 doi:10.1038/sj.jim.7000266 Received 11 February 2002/ Accepted in revised form 09 April 2002     

Differential affinities of <Emphasis Type="Italic">Erythrina cristagalli</Emphasis> lectin (ECL) toward monosaccharides and polyvalent mammalian structural units

Previous studies on the carbohydrate specificities of Erythrina cristagalli lectin (ECL) were mainly limited to analyzing the binding of oligo-antennary Galβ1→4GlcNAc (II). In this report, a wider range of recognition factors of ECL toward known mammalian ligands and glycans were examined by enzyme-linked lectinosorbent and inhibition assays, using natural polyvalent glycotopes, and a glycan array assay. From the results, it is shown that GalNAc was an active ligand, but its polyvalent structural units, in contrast to those of Gal, were poor inhibitors. Among soluble natural glycans tested for 50% molecular mass inhibition, Streptococcus pneumoniae type 14 capsular polysaccharide of polyvalent II was the most potent inhibitor; it was 2.1 × 10⁴, 3.9 × 10³ and 2.4 × 10³ more active than Gal, tri-antennary II and monomeric II, respectively. Most type II-containing glycoproteins were also potent inhibitors, indicating that special polyvalent II and Galβ1-related structures play critically important roles in lectin binding. Mapping all information available, it can be concluded that: [a] Galβ1→4GlcNAc (II) and some Galβ1-related oligosaccharides, rather than GalNAc-related oligosaccharides, are the core structures for lectin binding; [b] their polyvalent II forms within macromolecules are a potent recognition force for ECL, while II monomer and oligo-antennary II forms play only a limited role in binding; [c] the shape of the lectin binding domains may correspond to a cavity type with Galβ1→4GlcNAc as the core binding site with additional one to four sugars subsites, and is most complementary to a linear trisaccharide, Galβ1→4GlcNAcβ1→6Gal. These analyses should facilitate the understanding of the binding function of ECL. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.