Cascade Bio

🧬 Sucrase plays a critical role in carbohydrate metabolism, catalyzing the hydrolysis of sucrose into glucose and fructose. From substrate recognition to product release, each step of its mechanism enables sucrase to efficiently break down sucrose. For this week's post on our October Enzyme of the Month, let’s dive into how sucrase operates at a molecular level: 🔍 The first step in sucrase’s mechanism is substrate binding, driven by specific interactions between sucrose and conserved residues in the active site. Crystallographic studies have revealed that as sucrose enters the catalytic pocket, a tryptophan residue stabilizes the disaccharide through π-stacking interactions with the glucose moiety. Hydrogen bonds from asparagine and glutamine residues further align sucrose, locking it into an optimal position. 🧪 The catalytic step involves acid-base chemistry coordinated by active site residues. A glutamate residue acts as a general acid, protonating the glycosidic oxygen, which destabilizes the bond between glucose and fructose. Simultaneously, a histidine residue serves as a general base, activating a nearby water molecule for a nucleophilic attack on the glucose carbon. This step generates a covalent intermediate—a transient glucose-enzyme complex—stabilized through interactions with a tyrosine residue in the active site, which helps prevent premature product release. 🧫 Finally, product release occurs through hydrolysis of the covalent intermediate. A strategically positioned water molecule is activated by histidine, which facilitates the breakdown of the glucose-enzyme complex. The conformational flexibility of a loop structure near the active site allows the enzyme to reset after each cycle. This structural change improves turnover efficiency and ensures the active site can accommodate subsequent sucrose molecules without steric hindrance. 🔍 Each step in sucrase’s catalytic cycle, from substrate binding to product release, involves finely tuned molecular interactions that enhance efficiency. At the same time, understanding sucrase’s mechanism has paved the way for significant breakthroughs in both research and applied sciences. These discoveries will open new doors, demonstrating how fundamental enzyme mechanisms can lead to transformative applications across disciplines.

Today we are excited to announce that Cascade has a new scientific advisor - Doug Hattendorf. Doug brings over 20 years of enzyme experience from his lab days at Codexis to Zymergen where he led the 14 person Enzymology and Protein Engineering team to his Business Development efforts at Ginkgo. Check out the image below for a quote about why Doug is excited to collaborate with Cascade. He's already been instrumental in helping us as we develop our own enzymes - more on that soon!

🧬 Our October Enzyme of the Month, Sucrase, long studied for its role in carbohydrate metabolism, continues to be a subject of scientific investigation. Recent studies explore how this enzyme operates beyond digestion, unveiling new pathways and applications that could influence industrial processes, metabolic research, and therapeutic treatments. Let’s dive in: 🔬 At the University of Toronto, a recent study investigated the structural flexibility of sucrase under varying pH conditions. Researchers used cryo-electron microscopy to reveal how the enzyme's active site adapts to acidic and neutral environments, mimicking the pH variations in human intestines. Their findings suggest that sucrase can alter its catalytic efficiency based on environmental triggers, paving the way for tailored enzyme therapies that work across diverse gut conditions. 📊 Meanwhile, biochemists have also been studying sucrase's potential in addressing metabolic disorders. This research has focused on enhancing sucrase’s stability through site-directed mutagenesis, creating a variant that remains active longer in the gastrointestinal tract. By altering specific amino acid residues, they improved the enzyme’s resistance to degradation by proteases, making it more effective for enzyme replacement therapies targeting chronic digestive conditions. This discovery holds promise for new treatments targeting disorders like sucrase-isomaltase deficiency and could provide insights into developing enzyme supplements with extended shelf lives. 🧪 Sucrase has also been explored for its industrial potential beyond food science. Researchers demonstrated that engineered sucrase enzymes could be used to convert complex sugars into bioethanol efficiently. Their work opens doors for sustainable biofuel production, offering a greener and more economical alternatives. This cross-disciplinary approach highlights the enzyme’s potential in the renewable energy sector. 🌍 The expanding body of research on sucrase highlights the enzyme’s importance beyond textbook biochemistry. These advancements, spanning biofuel, therapeutics, and environmental sustainability, show how sucrase is driving breakthroughs across disciplines.

🍬 Sucrase, an enzyme that breaks down sucrose into glucose and fructose, has found its place not only in our digestive system but also across various industries. As an essential component of carbohydrate metabolism, its ability to hydrolyze glycosidic bonds has opened up a range of applications that go beyond the lab. In industrial settings, sucrase is harnessed to enhance sweetness, support fermentation processes, and facilitate therapeutic formulations, making it a versatile tool in biotechnology. Let’s dive in: 🧪 Sucrase is used in the production of sugars. By breaking down sucrose into its monosaccharides, glucose, and fructose, sucrase helps create sweeteners that are extensively used in food and beverage manufacturing. The enzymatic breakdown allows manufacturers to control the balance of sugars, optimizing flavor profiles while potentially reducing costs. 🍷 Another significant use of sucrase is in the fermentation industry, particularly in the production of alcoholic beverages. In wine-making and brewing, sucrase assists in breaking down residual sugars, providing a consistent supply of fermentable sugars for yeast. This enzymatic activity ensures efficient fermentation, leading to better yield and quality of the final product. By aiding in the conversion of complex sugars, sucrase contributes to the development of specific flavor profiles, which can be tailored to the desired characteristics of the beverage. 💊 Sucrase also plays a crucial role in the pharmaceutical industry, where it is utilized in the formulation of enzyme replacement therapies for individuals with congenital sucrase-isomaltase deficiency (CSID). CSID is a condition where the body lacks sucrase activity, leading to malabsorption and gastrointestinal issues when consuming sucrose. By incorporating sucrase into therapeutic formulations, it enables better digestion and absorption of sucrose, improving quality of life for affected individuals. The enzyme's stability and efficacy in such treatments demonstrate its adaptability and precision in addressing specific metabolic needs. 🔍 The industrial applications of sucrase highlight its importance beyond basic biochemistry, revealing its versatility in food production, fermentation, and therapeutic contexts. As we continue to explore the potential of enzymes like sucrase, their role in streamlining industrial processes and improving product formulations becomes increasingly evident. This enzyme serves as a reminder of how fundamental biochemical mechanisms can be repurposed for impactful innovations across multiple sectors. Tune in next week to learn more about our October Enzyme of the Month!

Congratulations to today's Nobel Prize winners David Baker, Demis Hassabis and John Jumper for their work developing the space of computational protein design and prediction. Proteins are beautiful structures, and admittedly, our simplified AI-generated images of enzymes don't do justice to all the twists, turns and folds that underpin nature's biochemical machinery. Thanks to the innovations of today's awardees, we understand this structural complexity better than ever before. We are still in the early days of understanding and utilizing proteins, and here at Cascade, we are excited to make proteins long-lasting, reliable and robust so they are well suited for a broad range of use cases across chemicals, food, pharma and more. #NobelPrize #Proteins #Enzymes #CascadeBio #Biomanufacturing

🎃 October is here, and with it comes Halloween—a perfect time to talk about Sucrase, an enzyme that helps break down table sugar (sucrose) into simpler sugars, glucose, and fructose. Sucrase plays a key role in our ability to enjoy sweet treats. Whether it's candy corn or chocolates, Sucrase ensures our body can digest these sugary delights properly. 🧬 The story of Sucrase stretches far beyond Halloween candy. Evolutionary biologists have traced the origins of Sucrase to ancient adaptations in mammals. As our ancestors' diets changed to include more carbohydrates, Sucrase became essential for survival, allowing us to efficiently process the energy found in sugars. 🧪 Discovered in the 19th century, Sucrase's isolation marked a milestone in understanding human digestion. Scientists identified its critical role in the small intestine, where it aids in breaking down sucrose from the foods we eat. Without Sucrase, sugar consumption would lead to digestive difficulties, linking its function directly to both nutrition and metabolic health. 🔬 On a molecular level, Sucrase is fascinating. It works by cleaving the glycosidic bond between glucose and fructose molecules. This seemingly simple action has complex implications for health, especially as we consider sugar intake in modern diets. Studying the mechanics of Sucrase helps us better understand how our bodies handle sugar and opens up discussions on how to process sugar more efficiently. ⚙️ All considered, Sucrase also holds exciting potential in enzyme engineering. Researchers are exploring ways to modify or enhance Sucrase to create more efficient versions for industrial applications. These engineered enzymes could be used to process sugars more effectively in food manufacturing or to develop treatments for digestive disorders where Sucrase activity is impaired. This field opens up new possibilities for biotechnology and nutrition science. With Sucrase as our October Enzyme of the Month, we’ll learn more about its applications, research, and mechanisms. 

Looking back on New York's Climate Week, I'm grateful for all the conversations I had with founders, potential investors, and other members of the ecosystem. Hopefully now, at least a few more people are excited about enzymes. Here are some highlights: On Monday, I reconnected with the Venture For ClimateTech team Jack Baron Devin Sandon Scott Meyers Cassie Schuttrumpf Jenny Leung and founders of Cohort 4 (Cascade was in Cohort 3). Congratulations to everyone for completing the program, and I enjoyed seeing a few companies also focused on cleaning up the chemicals industry. On Tuesday, I spent much of the day getting smarter on different methods of funding for capital-intensive businesses for when Cascade is operating at scale. Thanks to Enduring Planet, Hannah Friedman at Planeteer Capital and other sponsors for organizing an intensive session earlier in the day and thank you to Jack Fritzinger for bringing together start-ups and funders for First of a Kind (FOAK) pitches in the evening. On Wednesday, I spent time with my fellow synthetic biology start-ups at a couple events organized by Juniper Zero. I am always amazed at the different ways to use biology and the number of problems that can be solved with bio-based approaches. And finally, on Thursday, I spent nearly the whole day at Cascade Bio's table at Newlab's New Climate Futures event. I believe this was the first ever Cascade booth, so thank you to Newlab's Founder Fellowship program for giving us prime real estate during the showcase. I think the biggest questions I am still thinking about are: why is it still so difficult and expensive to move molecules around? Why do we need facilities costing hundreds of millions of dollars to do transformations that nature does for free? After this week, my belief in Cascade's vision for transforming how molecules are made—and its potential to address major climate challenges—is stronger than ever. Now it is time to get back to work.

In this edition of our Enzyme of the Month series, we’re taking a deep dive into the Eugenol biosynthesis cascade—the series of enzyme-driven reactions responsible for producing Eugenol, the warm and spicy compound found in cloves, cinnamon, and nutmeg. Each enzyme in this cascade shapes the molecular structure of Eugenol step by step. Let’s break down the key enzymes involved: 🌱 Phenylalanine Ammonia-Lyase (PAL) was not only our April Enzyme of the Month but is the first enzyme in the Eugenol pathway, responsible for converting the amino acid phenylalanine into cinnamic acid by removing an ammonia group. Structurally, PAL is a homotetramer, with each subunit containing an active site that catalyzes the deamination reaction. This enzyme plays a pivotal role in plant secondary metabolism, serving as a gatekeeper for phenylpropanoid biosynthesis. The reaction it catalyzes involves breaking the C–N bond in phenylalanine, forming a double bond between the α-carbon and the β-carbon, a critical first step in producing aromatic compounds like Eugenol. 🧪 Cinnamate-4-Hydroxylase (C4H) introduces a hydroxyl group at the 4-position of the cinnamic acid, converting it into p-coumaric acid. As a cytochrome P450 enzyme, C4H relies on electron transfer mechanisms to insert oxygen into the substrate, facilitated by a heme cofactor in its active site. The enzyme’s structural architecture includes a highly conserved P450 fold, optimized for substrate binding and precise hydroxylation. This step is critical for modifying the core phenylpropanoid skeleton, creating a versatile intermediate that can be further processed into various plant defense compounds, including Eugenol. 🔬Eugenol Synthase (EGS) is the final enzyme in the cascade, converting coniferyl alcohol into Eugenol through a reduction reaction. EGS belongs to the NADPH-dependent reductase family, utilizing NADPH to transfer electrons and reduce the double bond in the precursor molecule. Structurally, it is a dimer, with each subunit binding both the substrate and cofactor in a well-defined active site pocket. The enzyme’s mechanism involves protonation of the α,β-unsaturated bond, resulting in the formation of the characteristic hydroxyl group on Eugenol. This step finalizes the transformation into the spicy, aromatic compound we recognize in cloves, cinnamon, and other spices. 🌿 The enzymes in the Eugenol cascade highlight the precision of plant biochemistry, from the initial deamination of the phenylalanine to the final reduction that produces Eugenol. Each enzyme contributes not only to flavor but also to the plant’s natural defense mechanisms. And with that let’s thank these enzymes for all the fall vibes they add to our lives. Stay tuned next week for October’s enzyme! #EnzymaticFuture #CascadeBio #Eugenol #Biochemistry #EnzymeOfTheMonth #CytochromeP450 #PlantBiochemistry #EnzymeMechanism

Earlier this week the Cascade Bio team took a break from immobilizing enzymes to go search for treasure. Luckily we solved all the clues, found the blue crystal and escaped without using a single hint. Thank you Mission Escapes Denver for hosting us. Do you love enzymes and want to collaboratively solve challenges in the lab (and in escape rooms)? We are hiring: https://lnkd.in/esWFsh-C

🍂 The natural production of Eugenol involves a fascinating cascade of enzymatic reactions, our September Enzymes of the Month. While well-known for producing the warm, spiced flavor of clove, Eugenol's biosynthetic pathway has significant applications beyond flavor. Let’s dive into 3 key applications that rely on Eugenol’s enzymatic cascade: 💊 One important application of Eugenol's pathway is the development of pain-relieving and anti-inflammatory drugs. By tweaking enzymes like Eugenol synthase, researchers can optimize the production of bioactive derivatives. These derivatives are explored as alternatives to synthetic analgesics, offering a more natural route to pain management. Additionally, Eugenol's role in triggering reactive oxygen species (ROS) scavenging opens doors for antioxidant therapies, targeting oxidative stress-related diseases like arthritis and neurodegeneration. 🦠 Another promising application is in the realm of biotechnology, where synthetic biology harnesses the pathway for scalable, sustainable Eugenol production in engineered microbial systems. By integrating genes from Eugenol-producing plants into yeast or bacteria, scientists can produce Eugenol in large quantities without needing traditional plant sources. This microbial production method is particularly important for industries looking to reduce reliance on plant harvesting, ensuring a consistent supply for sectors like cosmetics, where Eugenol's antimicrobial and preservative properties are used in formulations for oral care, skincare, and personal hygiene products. 🌾 In agriculture, Eugenol's natural defensive properties against pests make the manipulation of its pathway highly relevant for developing bio-pesticides. The cascade involved in its synthesis allows for fine-tuning the compound's potency and targeting specific pests without harming beneficial insects or plants. This application is part of a broader movement towards natural, environmentally friendly pest control methods, providing an alternative to synthetic chemicals. The ability to enhance or modify this pathway to create pest-resistant crops could have substantial implications for sustainable farming practices globally. 🔗 The enzymatic pathway for Eugenol production showcases how fine biochemical tuning can lead to diverse, impactful applications. Whether in medicine, biotechnology, or agriculture, the exploration and manipulation of this cascade continue to push the boundaries of what natural compounds can do, revealing new opportunities for both scientific innovation and practical solutions. #EnzymaticFuture #CascadeBio #CascadeBiocatalysts #Eugenol #PumpkinSpice #PSL #Biochemistry #EnzymeOfTheMonth #Research #Antioxidants