From lab bench to formulated ingredient: Characterization, production, and commercialization of human milk oligosaccharides

Walsh, Clodagh; Lane, Jonathan A.; Sinderen, Douwe van; Hickey, Rita M.

doi:10.1016/j.jff.2020.104052

Cited by 48 publications

(51 citation statements)

References 140 publications

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“…Human milk oligosaccharides (HMOs) furnish breast-fed infants with a number of health benefits. Their unique composition, differing from other mammal’s milk, drives active research to synthesize and produce the main HMO structures as supplements for infant formula milks (for recent reviews, see [ 1 , 2 , 3 , 4 , 5 ]). Various beneficial effects are critically important for early development and infant health, functioning as prebiotics and antimicrobial agents as well as exerting immunomodulation effects [ 6 , 7 ].…”

Section: Introductionmentioning

confidence: 99%

Transglycosylation Activity of Engineered Bifidobacterium Lacto-N-Biosidase Mutants at Donor Subsites for Lacto-N-Tetraose Synthesis

Castejón-Vilatersana

Faijes

Planas

2021

IJMS

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The health benefits of human milk oligosaccharides (HMOs) make them attractive targets as supplements for infant formula milks. However, HMO synthesis is still challenging and only two HMOs have been marketed. Engineering glycoside hydrolases into transglycosylases may provide biocatalytic routes to the synthesis of complex oligosaccharides. Lacto-N-biosidase from Bifidobacterium bifidum (LnbB) is a GH20 enzyme present in the gut microbiota of breast-fed infants that hydrolyzes lacto-N-tetraose (LNT), the core structure of the most abundant type I HMOs. Here we report a mutational study in the donor subsites of the substrate binding cleft with the aim of reducing hydrolytic activity and conferring transglycosylation activity for the synthesis of LNT from p-nitrophenyl β-lacto-N-bioside and lactose. As compared with the wt enzyme with negligible transglycosylation activity, mutants with residual hydrolase activity within 0.05% to 1.6% of the wild-type enzyme result in transglycosylating enzymes with LNT yields in the range of 10-30%. Mutations of Trp394, located in subsite -1 next to the catalytic residues, have a large impact on the transglycosylation/hydrolysis ratio, with W394F being the best mutant as a biocatalyst producing LNT at 32% yield. It is the first reported transglycosylating LnbB enzyme variant, amenable to further engineering for practical enzymatic synthesis of LNT.

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Section: Introductionmentioning

confidence: 99%

Transglycosylation Activity of Engineered Bifidobacterium Lacto-N-Biosidase Mutants at Donor Subsites for Lacto-N-Tetraose Synthesis

Castejón-Vilatersana

Faijes

Planas

2021

IJMS

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“…Chemoenzymatic techniques, together with microbial metabolic engineering processes, are currently used to produce HMO in sufficient quantities to be able to address the design of functional and nutraceutical foods, in addition to preclinical and clinical studies to evaluate them [ 94 , 95 ]. However, to emulate the biological function of HMOs, structure–function relationships must be taken into account, which is a major challenge for the biotechnology industry [ 96 ]. Although the synthesized HMOs show a structural identity with the natural ones, the introduction of these ingredients requires the evaluation and approval of the different administrations in the world.…”

Section: Human Milk Oligosaccharides (Hmos)mentioning

confidence: 99%

“…There are several of these synthetic HMOs already approved on the market and some of them (2′-fucosillactose (2′-FL) and lacto-N-neotetraose (LNnT)) incorporated in commercialized infant milk, with good results in terms of safety, tolerance, and intestinal benefits in intervention studies [ 97 , 98 , 99 ]. They are also beginning to be incorporated into products for a wider audience [ 96 ]. More HMOs are expected to be added to the international market as the different administrations authorize them.…”

Section: Human Milk Oligosaccharides (Hmos)mentioning

confidence: 99%

Breast Milk: A Source of Functional Compounds with Potential Application in Nutrition and Therapy

et al. 2021

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Breast milk is an unbeatable food that covers all the nutritional requirements of an infant in its different stages of growth up to six months after birth. In addition, breastfeeding benefits both maternal and child health. Increasing knowledge has been acquired regarding the composition of breast milk. Epidemiological studies and epigenetics allow us to understand the possible lifelong effects of breastfeeding. In this review we have compiled some of the components with clear functional activity that are present in human milk and the processes through which they promote infant development and maturation as well as modulate immunity. Milk fat globule membrane, proteins, oligosaccharides, growth factors, milk exosomes, or microorganisms are functional components to use in infant formulas, any other food products, nutritional supplements, nutraceuticals, or even for the development of new clinical therapies. The clinical evaluation of these compounds and their commercial exploitation are limited by the difficulty of isolating and producing them on an adequate scale. In this work we focus on the compounds produced using milk components from other species such as bovine, transgenic cattle capable of expressing components of human breast milk or microbial culture engineering.

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“…For example, differences in gut microbiota composition between breastfed and formulafed infants are largely attributable to the presence of human milk oligosaccharides (HMOs) in breast milk [13]. HMOs are complex sugars with unique structural diversity and are known to positively influence infant health [14,15]. Recent progress in HMO manufacturing and chemoenzymatic synthesis has increased the availability of these oligosaccharides, thereby facilitating more in-depth studies of their function and the effects of their supplementation in infant formulae.…”

Section: Introductionmentioning

confidence: 99%

“…Recent progress in HMO manufacturing and chemoenzymatic synthesis has increased the availability of these oligosaccharides, thereby facilitating more in-depth studies of their function and the effects of their supplementation in infant formulae. Notably, a few studies in infants have successfully confirmed the safety and efficacy of HMO-fortified infant formulae [14,16], supporting the regulatory approval of two major HMOs: 2 -fucosyllactose and lacto-N-neotetraose [15]. However, data regarding the effects of HMOs on bacterial adherence are scarce.…”

Section: Introductionmentioning

confidence: 99%

Identification of Synbiotics Conducive to Probiotics Adherence to Intestinal Mucosa Using an In Vitro Caco-2 and HT29-MTX Cell Model

et al. 2021

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The ability of bacteria to adhere to the intestinal mucosa is a critical property necessary for the long-term colonization of the intestinal tract. This ability can be highly sensitive to the presence of prebiotics. However, limited data are available in this respect for beneficial bacteria such as probiotics or resident gut microbiota. We previously demonstrated that the presence of prebiotics may decrease adherence in several pre- and probiotic combinations. Thus, characterizing the interactions between numerous combinations involving different classes of pre- and probiotics can be crucial in identifying new synbiotics. Accordingly, here, we extend our prior analyses to evaluate the adhesion of five lactobacilli, six bifidobacteria, and one probiotic Escherichia coli strains, as commercial probiotics or promising probiotic candidates, together with the cariogenic Bifidobacterium dentium strain. As an in vitro intestinal mucosa model, Caco-2 and mucin-secreting HT29-MTX cells were co-cultured at 9:1 in the presence or absence of prebiotics. Commercial inulin-type fructooligosaccharide prebiotics Orafti® GR, Orafti® P95, and galactooligosaccharide-based prebiotic formula Vivinal®, including purified human milk oligosaccharides (HMOs) were added into the cultivation media as the sole sugar source (2.5% each). Adherence was tested using microtiter plates and was evaluated as the percentage of fluorescently labeled bacteria present in the wells after three washes. Consistent prebiotics-mediated enhanced adherence was observed only for the commercial probiotic strain E. coli O83. For the remaining strains, the presence of HMO or prebiotics Orafti® P95 or Orafti® GR decreased adherence, reaching statistical significance (p < 0.05) for three of out of eight (HMO) or five of out of 11 strains tested, respectively. Conversely, Vivinal® enhanced adhesion in six out of the 12 strains tested, and notably, it significantly attenuated the adherence of the cariogenic Bifidobacterium dentium Culture Collection of Dairy Microorganisms (CCDM) 318. To our knowledge, this represents the first report on the influence of commercial prebiotics and HMOs on the adhesion of the cariogenic Bifidobacterium sp. Vivinal® seems to be a promising prebiotic to be used in the formulation of synbiotics, supporting the adhesion of a wide range of probiotics, especially the strains B. bifidum BBV and BBM and the probiotic Escherichia coli O83.

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From lab bench to formulated ingredient: Characterization, production, and commercialization of human milk oligosaccharides

Cited by 48 publications

References 140 publications

Transglycosylation Activity of Engineered Bifidobacterium Lacto-N-Biosidase Mutants at Donor Subsites for Lacto-N-Tetraose Synthesis

Transglycosylation Activity of Engineered Bifidobacterium Lacto-N-Biosidase Mutants at Donor Subsites for Lacto-N-Tetraose Synthesis

Breast Milk: A Source of Functional Compounds with Potential Application in Nutrition and Therapy

Identification of Synbiotics Conducive to Probiotics Adherence to Intestinal Mucosa Using an In Vitro Caco-2 and HT29-MTX Cell Model

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