Direct Acid Methylation for Extraction of Fatty Acid Content from Microalgae Cells

Direct Acid Methylation for Extraction of Fatty Acid Content from Microalgae CellsDirect Acid Methylation for Extraction of Fatty Acid Content from Microalgae Cells

By: Benjamin D. Frigo-Vaz, Ping Wang

Abstract:
Direct acid methylation was examined as a means for both analysis of fatty acid content in microalgal cells and biodiesel production without pretreatment. Microalgal cells of Chlamydomonas reinhardtii and Dunaliella tertiolecta were prepared and examined. It appeared that direct acid methylation extracted higher fatty acid content than the solvent-based Soxhlet extraction process. It also revealed that the latter was prone to extract a significant amount of nonlipid hydrophobic impurities, including hydrophobic proteins and phytol-type compounds, while direct methylation produces essentially pure ester product. This work demonstrates that direct acid methylation provides superior fatty acid extraction, promising an efficient process for either quantification of lipid content or production of biodiesel.

Link to Applied Biochemistry and Biotechnology Journal for book.

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Agua Potable de la humedad del aire, Water from the humidity in the air

Clean water, producing water from the air humidity

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Nationalism is Modern Tribalism… and still as brutal!

The rising conflicting between Russia and Ukraine reminds me of how much I blame nationalism as a fundamental evil of humanity. Likely born long before we even came down from the trees, humans have an ingrained instinctive desire to form groups or “tribes” and to distinguish each other as “us” and “them”. This instinct to tribalize is the fundamental cause of all wars, all racism and all conflicts between groups, for if we did not create these groups (and I emphasis “create” here for all of these categories are mostly or completely imaginary.) none of those conflicts would occur! Mind you this instinct brought humanity great benefit throughout most of our evolution, at least for the victors.. the loser got murder, rape and enslavement, and if they had any proclavities counter to tribalism those got skimmed off the gene pool very quickly.

Many today still think of our old tribal days a human’s good-old-days when we living in “harmony” with nature and some kind of utopian anarchism. That harmony was achieved by brutish existence where natural mechanism of population control, disease, famine, and tribal warfare, kept our populations stable by force of pain, suffering and carnal murder! Don’t believe me? Read the works of Steven Pinker and his kind for starters, begin by watch his TED talk

Thankfully human’s can be more then impulsive animals, we have created though acquired knowledge, reasoning and ethics, overroad nature and achieve levels of happiness and passivity in the last few thousand years that would not have been naturally possible. Unfortunately our knowledge and social systems have evolved at a lamarckian pace, thousands of times the speed of the Darwinian evolution of our genetics and as such we still retain all these vile instincts that are counter-productive to our social progress and even logical thought.

Back to Urkaine and Russia. Russia declares it protecting Russian people and “Russian speakers” from Ukrainians, claiming (with no evidence) that they are under attack. So lets ask some rhetorical questions: Who being protected? People, from other people supposedly… why? Because one of them are like us (or us) and the others are “the others”, the impulse is clear, we must crush and take from the other, they are evil, we are good. It not hard to see how silly this is if your willing to strip the modern facade of nationalism off of its tribalist core. The Ukrainians likewise want a Ukrainian for Ukrainians and don’t want to give up any land that is “theirs” to “Other” people, that being the Russians. This very concept of the nation, of needing to fight for your “nation” against other “nations” is as silly and stupid as fighting for your clan, tribe or race. Heck at least the latter are based on something real like genetic relationship: only humans can create completely imaginary groupings based on concepts alone… and then kill each other over these groupings just like chimps over territories.

The concepts that make up a nation, its language, its culture exist only in the heads of people, even the sounds and symbols humans create to try to manifest something physical for this culture only has meaning to those people in their head… and yet we can kill each other over this? I might understand if this was a war over the kind of government wanted, a war over what rights people should have, but no this is a war fundamentally over what language people speak and what color flag they want. Humanity has progressed so far to bring the clan to the tribe, the tribe to the kingdom, the kingdom to the state and the state to the nation, now even a united nations exist (though powerless), it not hard to see that nations will eventually give way to a singular humanity, but for that day to come would we not need a singular culture and language or can we evolve enough to accept, tolerate and even celebrate such variety? No, culture adds beauty to life and thought, it can remain, the problem is not culture it is the ancient tribalist urge to separate us and them. We can choose to distinguish on race, we can choose to distinguish on religion, we can choose to distinguish on country, or we can choose not to and end all the silly blood shedding, it requires only a minor bit more force of will then we have already made to bring society this far from time of tribalism.

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The Gender Equality Test

Modern Feminism, Egalitarianism and Humanism all converge on the ideal gender equality. All these agree on gender equality being a goal, but few adherents have more than a vague understanding of what gender equality is. For that I have a simple test:

1. Take any situation, lets say for example: “Women slaps her husband for cheating on her”

2. Give it a moral value: “how right or wrong is this situation?”

3. Keep everything the same, but reverse the genders: “Man slaps his wife for cheating on him”

4. Give this second situation a moral value.

5. Compare the moral value of the two situations. If they are the same to you, if they are equally right or equally wrong, then that is gender equal. If they are not the same to you then you don’t believe in gender equality.

It is really that simple!

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The Future is Automated (Part 1)

The standard Anti-Luddite argument is that as jobs are taken by automation in one field people start working in another field. This is reasonably backed by history but there is no evidence to assume it will hold true indefinitely and significant evidence it is now not holding true anymore.

The introduction of the steam engine and industrial machines first affected agriculture. Automated thrashers, cotton pickers, combines rendering most agriculture jobs obsolete decades to centuries ago. Agriculture employed (or enslaved) over 70% of the American population back in 1800
Today it only accounts for 2% of the labour force, yet produces enough food for over 60 times the 1800 population (not including food relief or sales to other countries around the world). So if agriculture went from 70% to 2%, all the workers had to go somewhere and they did: into manufacturing. But by 1950 the so called golden age of US manufacturing 35% of the labor force was working in factories, today that number is 9%. Yet US manufacturing has only risen in production and is still the worlds largest manufacturing center.

Today 80% of the labour force in the USA is in the “Service sector”, but automation is making inroads in that as well. In both Federico Pistono book “Robots will Steal Your Jobs but that’s OK” and Martin Ford book “The Lights in the Tunnel: Automation, Accelerating Technology and the Economy of the Future” they provide a break down of the most common jobs in the USA today. For example cashiers, 2nd most common job is 2.24% of all American jobs, but now self-checkout lanes are being installed and can cut down the number of cashiers by at least 75%. Truck drivers (the most common job in the USA) are 2.61% but automated trucks will likely start taking those jobs as well. Even if a truck driver is required in the automated truck as backup the driver can now sleep behind the wheel while the truck drives to its destination without having to stop so the driver can sleep, allowing fewer drivers to do more deliveries. “No these people will move on to new fields” so the anti-Luddite claims. What new fields are there? In the list of jobs from most common to least you have to go down to 34th on the list to get to a career that has not existed more then 70 years ago: Computer Software Engineer at 0.74% of the job market.

Well maybe people working in service today could get work in intellectual and skilled labour fields. Unfortunately there are in fact a plethora of inroads automation is making in those fields as well: pharmacist replaced by robots, software that is replacing paralegals and lawyers in document review, software that can write journalistic sports and basic business reports, image recognition software that replaces radiologist. It is argued that these things augment and assist people in these fields but if automation helps one person do more of a task then would it not mean that now less people need to do those tasks?

Another way to look at this is as a ratio of number of products produces divided by number of people needed to produce said products. Over the centuries this ratio has been going up in general for most products due to automation, but demand went up to match. As a result everyone had been able to get a job even with more and more automation because automation just means more products per producing person. Everyone can be productive because productivity was matched by demand, but can this stay true forever? Every product requires some amount of physical resources and energy to make as well as demand from a limited number of end users. Lets look at energy (as material resources are just a matter of how much energy is needed to to extract, refine and move them): For the last 3 centuries world energy consumption has been growing relatively continuously by ~3% per year. Right now it is at 15×10^12 watts (15 Terawatts), at 3% growth it would be at 150 TW by 2091 10, times as much in 78 years! By 2345 our energy consumption would be equal to ALL the energy in sunlight hitting the earth (2.7×10^17 W)! Even if we could produce all that power by nuclear fusion or something the waste heat would exceed solar heating of the planet! The surface temperature of the earth would raise to boiling, literally boiling in just a few years from our activity alone! If we keep projecting 3% growth by the year 3058 humanities energy consumption would exceed the energy output of the entire sun (3.8×10^26 W)! And by the year 3760 humanities energy consumption would exceed the output of every single star in the galaxy (3.8×10^35 W)! Hence why there is no such thing as sustainable growth, it is PHYSICALLY IMPOSSIBLE! Pistono does a good job explaining this exponential growth problem but fails to really connect it to automation, I will now do that.

Thankfully world total energy consumption is starting to stagnate (actually falling in 2009!) and consistently falling in many developed countries. World population growth is slowing (and will hopefully plateau this century, and thus save future generations from mass starvation or heat death as mentioned above) and some developed countries even have negative population growth. But though that may save us from physical limits of energy and matter its not good for production. How are we to produce more products if there is not enough power and raw materials to make them and consumers to buy them? If automation keeps growing the ratio but the number of products can’t keep going up, then number of people needed to make them must go down.

Imagine for a moment a restaurant where the owner gets a machine to replace most if not all the workers, anyone that has worked at a McDonalds as a teenager can tell you that it would not be very hard to replace the assembly line of human burger makers with a machines that cooks the meat paddies, drops them on the bread buns, drops down pickles, lettuce, tomatoes and squirt of condiments. Now lets say the workers ran off to start restaurants of their own using the same model. Thus multiplying the number of restaurants, everyone gets a high paying job but would there be enough consumers for them all? Agriculture has experience several fold increases in products to producers ratio but the number of consumers has increased to match it so the ratio has increased much less if we look at it from a products to consumer ratio rather the products to producers ratio. Still it increased enough that only a tiny fraction needs to work in that field now. As is Americans are eating way too much, many are morbidly obese and dying of diabetes and heart disease: we would need to institute real vomitoriums to get people to “consume” more food! But this scenario does not just play out with foods. Imagine cellphone makers replace all there labors with machines, and that magically those labours got the education to design new cellphones and manage new cellphone companies, with a production to R&D ratio of greater then 100:1, could we all stand to buy 100 cell phones to keep all those new cell phone companies afloat? This logic can be extended to all products all people even the filthy rich don’t buy incredible amounts of stuff like a kleptomaniac on speed! Everyone’s (unless mentally disturbed and/or on drugs) consumption will become satiated with enough supply. And because population growth is slowing and will hopefully plateau production will eventual reach the limit of consumer saturation.

Now some may argue that automation can’t take ALL jobs, well at present and for the near future that seems true. Most automation is done by at best Weak Artificial Intelligence: very “smart” at a specific task and completely incompetent at anything else. The IBM’s Watson supercomputer and DeepQA program can and has trumped the best humans at answering even cryptic questions from memorized knowledge, it certainly is smart at that and will likely replaced tech support workers and research aids, but it can’t create new new data, or assemble a product or drive a car. Other WAIs though can do each one of those task from Self-evolving software that discovers new technologiess, to image recognizing factor robots, to self driving cars. Yet WAIs can only do those tasks they are specifically programmed and built for: they can’t think about or are even “aware” of anything else, they have no consciousness. A Strong Artificial Intelligence that could solve or adapt to any problem a human can and/or could qualify as conscious is likely decades away, and some theologians and philosophers say its impossible because of some supposed mystical and supernatural property of the human brain. SAI though is not nessassary: machines don’t need to take all jobs, just enough, fast enough, its simply a matter of automation taking jobs and incomes faster then they can be replaced. Imagine if say 30% of job force are forever safe from automation, that would mean 2/3 of the population would be unemployed and unemployable! If we assume everyone could move to doing what jobs that are left and that somehow those remaining fields could grow to fill everyone’s employment needs its likely most of those jobs that are left would not be high paying or desirable, as is the service sector switch for most workers has had detrimental affects on the economy.

Imagine machines will soon replace most, but not all service jobs, what jobs are left? Maintaining the machines? Sure but only a few people are needed to do that verse all the number of people who did those jobs the machines replaced. Ok how about jobs as engineers, biochemist, inventors… but those require extensive educations, above average intelligence and creativity. Ok how about artist and craftmen? Again you the need for creativity, rarefied skill and talent, not enough people will be around to buy all those arts and crafts. There won’t be more people having plumbing problems and needing more plumbers, having electrical problems and needing more electricians, heck if you have a computer problem these day you usually contact a machine first that can answer some of your problems rather then a paid specialist. Even if automation does not replace a job outright it replaces enough parts of that job to make the worker more efficient and effective, and without more demand to match the increase productivity there will need to be less workers.

If automation enters an industry but the industry can’t grow to match (because of physical limitation at most), then its work force must shrink, we have the historical drop of agriculture and manufacture jobs as proof of that. The profits that was taken to pay wages now goes up to the owners of the businesses, and the unemployed workers must compete in a shrinking job market, this would cause incomes to drop, middle class to shrink and become poor, the poor grow poorer and the rich to grow richer. That is exactly what is happening today! Stock markets and capital gains are higher then they have ever been before, yet unemployment is up, people are getting lower and lower paying part time jobs. The middle class is shrinking into poverty.

“The top 10 percent of Americans have experienced rapid income growth over the last 40 years, but the bottom 90 percent haven’t been so lucky. In fact, average income rose just $59 from 1966 to 2011 for the bottom 90 percent once those incomes were adjusted for inflation.”

That’s right we have not moved our income up for the common American for nearly 50 years now! Yet US farm production has gone up, US manufacturing production has gone up, capital has grown. Most Americans have moved to poorer paying part-time dead-end service jobs, that most of them hate!

Much of this is blamed on outsourcing now, a problem only in that its giving someone else “your” job. Outsourcing is a problem for the working population of developed nations, it drives salaries down to the lowest common denominators of the world. But truth be known machines are now competing with developing country’s cheap practically slave labour! For example Foxcon the Taiwanese electronic giant that assembles all Apple products of all things is beginning to to automate. It is all a matter of when will machines cost less then a practical slave in a undeveloped country. Also unskilled impoverished labours can’t compete in quality verse the precision of automatons, nor will automatons commit suicide or demand pay raises or human rights, machines as long as they are design not to will never feel, never want, never need.

To truly determine if automation is a greater problem then outsources simply compare the product to worker ratio there to here:

“if 10% of China’s electronics production was moved to the U.S., China would lose 300,000 jobs. Yet, just 40,000 new jobs would be created in the U.S. Put another way, if all of China’s manufacturing output was magically transported to the U.S. tomorrow, the U.S. unemployment rate would decline by only 2.75 percentage points after accounting for the effects of automation.”

This can’t continue for long. If more products could be produced maybe their price would drop and the ever poorer people could still buy them but as I covered before there are fundamental limits in energy and materials that we appear to already be hitting that limit production. As the people get poorer they can’t buy the products anymore. So demand will drop, and that means the rich will start to see profits drop. In fact the economy of individual nations and thus every nation because of globalization will become increasingly unstable and fall into recessions one after another. Oscillating wildly or possibly even entering depressions until solutions are implement.

So what are the solutions, what can you personally do to survive the present yet still mostly unnoticed “Autopocolyse”? I will talk about the solutions in my next blog, some of which could lead to a future as wonderful as today would be to a black slave of 1800 America, but for now, I need you to understand the gravity of the problem, that it is happening now and its only going to get worse.

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RuBisCO: Natures Most Horrible Mistake

No I’m not making up words. RuBisCO is the arguably most important enzyme on the planet, its function is the first step in the capturing of carbon dioxide in plants in order to make biological matter, ergo it is where all the work of photosynthesis gets done, and that no exaggeration because it is so inefficient and its fidelity so poor that the single greatest efficiency loss in photosynthesis is here!

RuBisCO is so painfully slow, garbing only a few carbon dioxide molecules per second that plants have to produce incredible amounts of this enzyme making it quite possibly the most common protein by mass on the planet! In some plants up to 50% of their protein mass is RuBisCO! Its fidelity is such that it will capture oxygen instead of carbon dioxide undergo a energy losing process called photorespiration in which substrates are broken down and energy must be spent simply to repair the substrates, this happens at least 1/4 of the time!

Plants have evolved rather hair-brain ways of trying to improve the efficiency and fidelity of RuBisCO. Take C4 plants for example which capture CO2 by a completely different pathway, only to pump the capture CO2 deeper inside the leaf, away from oxygen and then release the CO2 in concentration around RuBisCO. This process is energy wasteful but worth it for the gain in efficiency and fidelity of RuBisCO. The only problem is dear god why not just cut RuBisCO out all together and use the CO2 captured by a faster more efficient pathway!

Evolution is not a smart process. I remember my bachelor degree days when I was making a Best-Fit Algorithm in MatLab, the algorithm would search through 12 dimensions of scalar and vectored data, picking points at random and asking how well its results fit the data, and then repeating around points that had had good results. Often the program would get stuck around localized mimima: areas where it could not make a better fit or get out of but was still off from the true answer. I relies evolution works like this: it finds answers at random through mutation and those answers may be far from the best possible but the mutations required to find better solutions are so unlikely, so out of range, that it gets stuck optimizing a poor solution rather then finding a better one. This is what I think RuBisCO is, an optimized poor solution that biology is stuck utilizing. This is why C4 plants have their hair-brain work around because to develop a completely alternate pathway would require so many new enzymatic steps that evolutionarily its too unlikely to happen and be competitive off the bat, hence the half-way pathway ‘dump it back on RuBisCO’ solution.

There is significant biotechnological work focused on genetic engineering a more efficient and accurate RuBisCO but so far they have not made significant progress, I don’t think they will, I think the enzyme has been optimized as best it can for each plant. The more they increase the rate of reaction the more they reduce the fidelity and increase photorespiration, and vis versa.

Why did biology get stuck with this pathetic enzyme? Well if you look at it from the fact when photosythesis evolved 3.5 billion years ago carbon dioxide concentrations in atmosphere were thousands of times greater then today and oxygen concentrations were non-existent, back then RuBisCO was probably the best photosynthetic pathway around and plants got stuck in that pathway to this day despite how inadequate RuBisCO is for today’s atmosphere. As a result our planets total carriage capacity is held down by a simple enzyme with a silly name.

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